Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2

Sung-Bau Lee, Chung-Fan Lee, Derick S C Ou, Kalpana Dulal, Liang-Hao Chang, Chen-Han Ma, Chien-Fu Huang, Hua Zhu, Young-Sun Lin, Li-Jung Juan

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Chromatin assembly factor 1 (CAF1) consisting of p150, p60 and p48 is known to assemble histones onto newly synthesized DNA and thus maintain the chromatin structure. Here, we show that CAF1 expression was induced in human cytomegalovirus (HCMV)-infected cells, concomitantly with global chromatin decondensation. This apparent conflict was thought to result, in part, from CAF1 mislocalization to compartments of HCMV DNA synthesis through binding of its largest subunit p150 to viral immediate-early protein 2 (IE2). p150 interaction with p60 and IE2 facilitated HCMV DNA synthesis. The IE2Q548R mutation, previously reported to result in impaired HCMV growth with unknown mechanism, disrupted IE2/p150 and IE2/histones association in our study. Moreover, IE2 interaction with histones partly depends on p150, and the HCMV-induced chromatin decondensation was reduced in cells ectopically expressing the p150 mutant defective in IE2 binding. These results not only indicate that CAF1 was hijacked by IE2 to facilitate the replication of the HCMV genome, suggesting chromatin assembly plays an important role in herpesviral DNA synthesis, but also provide a model of the virus-induced chromatin instability through CAF1. © 2011 IBCB, SIBS, CAS All rights reserved.
Original languageEnglish
Pages (from-to)1230-1247
Number of pages18
JournalCell Research
Volume21
Issue number8
DOIs
Publication statusPublished - 2011
Externally publishedYes

Fingerprint

Chromatin Assembly Factor-1
Immediate-Early Proteins
Cytomegalovirus
Chromatin
Histones
DNA
Chromatin Assembly and Disassembly
Viral Proteins
Human Genome
cytomegalovirus immediate-early proteins
Protein Binding
Viruses

Keywords

  • chromatin assembly factor 1
  • chromatin structure
  • human cytomegalovirus
  • immediate-early protein 2
  • histone
  • IE2 protein, Cytomegalovirus
  • immediate early protein
  • transactivator protein
  • virus DNA
  • amino acid substitution
  • article
  • cell line
  • chromatin
  • chromatin assembly and disassembly
  • Cytomegalovirus
  • genetics
  • host pathogen interaction
  • human
  • metabolism
  • protein binding
  • protein subunit
  • virus replication
  • Amino Acid Substitution
  • Cell Line
  • Chromatin
  • Chromatin Assembly and Disassembly
  • Chromatin Assembly Factor-1
  • DNA, Viral
  • Histones
  • Host-Pathogen Interactions
  • Humans
  • Immediate-Early Proteins
  • Protein Binding
  • Protein Subunits
  • Trans-Activators
  • Virus Replication
  • Human herpesvirus 5

Cite this

Lee, S-B., Lee, C-F., Ou, D. S. C., Dulal, K., Chang, L-H., Ma, C-H., ... Juan, L-J. (2011). Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2. Cell Research, 21(8), 1230-1247. https://doi.org/10.1038/cr.2011.53

Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2. / Lee, Sung-Bau; Lee, Chung-Fan; Ou, Derick S C; Dulal, Kalpana; Chang, Liang-Hao; Ma, Chen-Han; Huang, Chien-Fu; Zhu, Hua; Lin, Young-Sun; Juan, Li-Jung.

In: Cell Research, Vol. 21, No. 8, 2011, p. 1230-1247.

Research output: Contribution to journalArticle

Lee, S-B, Lee, C-F, Ou, DSC, Dulal, K, Chang, L-H, Ma, C-H, Huang, C-F, Zhu, H, Lin, Y-S & Juan, L-J 2011, 'Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2', Cell Research, vol. 21, no. 8, pp. 1230-1247. https://doi.org/10.1038/cr.2011.53
Lee, Sung-Bau ; Lee, Chung-Fan ; Ou, Derick S C ; Dulal, Kalpana ; Chang, Liang-Hao ; Ma, Chen-Han ; Huang, Chien-Fu ; Zhu, Hua ; Lin, Young-Sun ; Juan, Li-Jung. / Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2. In: Cell Research. 2011 ; Vol. 21, No. 8. pp. 1230-1247.
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title = "Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2",
abstract = "Chromatin assembly factor 1 (CAF1) consisting of p150, p60 and p48 is known to assemble histones onto newly synthesized DNA and thus maintain the chromatin structure. Here, we show that CAF1 expression was induced in human cytomegalovirus (HCMV)-infected cells, concomitantly with global chromatin decondensation. This apparent conflict was thought to result, in part, from CAF1 mislocalization to compartments of HCMV DNA synthesis through binding of its largest subunit p150 to viral immediate-early protein 2 (IE2). p150 interaction with p60 and IE2 facilitated HCMV DNA synthesis. The IE2Q548R mutation, previously reported to result in impaired HCMV growth with unknown mechanism, disrupted IE2/p150 and IE2/histones association in our study. Moreover, IE2 interaction with histones partly depends on p150, and the HCMV-induced chromatin decondensation was reduced in cells ectopically expressing the p150 mutant defective in IE2 binding. These results not only indicate that CAF1 was hijacked by IE2 to facilitate the replication of the HCMV genome, suggesting chromatin assembly plays an important role in herpesviral DNA synthesis, but also provide a model of the virus-induced chromatin instability through CAF1. {\circledC} 2011 IBCB, SIBS, CAS All rights reserved.",
keywords = "chromatin assembly factor 1, chromatin structure, human cytomegalovirus, immediate-early protein 2, histone, IE2 protein, Cytomegalovirus, immediate early protein, transactivator protein, virus DNA, amino acid substitution, article, cell line, chromatin, chromatin assembly and disassembly, Cytomegalovirus, genetics, host pathogen interaction, human, metabolism, protein binding, protein subunit, virus replication, Amino Acid Substitution, Cell Line, Chromatin, Chromatin Assembly and Disassembly, Chromatin Assembly Factor-1, DNA, Viral, Histones, Host-Pathogen Interactions, Humans, Immediate-Early Proteins, Protein Binding, Protein Subunits, Trans-Activators, Virus Replication, Human herpesvirus 5",
author = "Sung-Bau Lee and Chung-Fan Lee and Ou, {Derick S C} and Kalpana Dulal and Liang-Hao Chang and Chen-Han Ma and Chien-Fu Huang and Hua Zhu and Young-Sun Lin and Li-Jung Juan",
note = "被引用次數:4 Export Date: 28 March 2016 通訊地址: Juan, L.-J.; Genomics Research Center, Academia Sinica, Taipei 115, Taiwan; 電子郵件: ljjuan@gate.sinica.edu.tw 化學物質/CAS: histone, 9062-68-4; Chromatin; Chromatin Assembly Factor-1; DNA, Viral; Histones; IE2 protein, Cytomegalovirus; Immediate-Early Proteins; Protein Subunits; Trans-Activators 參考文獻: Smith, S., Stillman, B., Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro (1989) Cell, 58 (1), pp. 15-25. , DOI 10.1016/0092-8674(89)90398-X; Li, Q., Zhou, H., Wurtele, H., Davies, B., Horazdovsky, B., Verreault, A., Zhang, Z., Acetylation of histone H3 lysine 56 regulates replication-coupled nucleosome assembly (2008) Cell, 134 (2), pp. 244-255. , DOI 10.1016/j.cell.2008.06.018, PII S0092867408007708; Takami, Y., Ono, T., Fukagawa, T., Shibahara, K.-I., Nakayama, T., Essential role of chromatin assembly factor-1-mediated rapid nucleosome assembly for DNA replication and cell division in vertebrate cells (2007) Molecular Biology of the Cell, 18 (1), pp. 129-141. , DOI 10.1091/mbc.E06-05-0426; Corpet, A., Almouzni, G., Making copies of chromatin: The challenge of nucleosomal organization and epigenetic information (2009) Trends Cell Biol, 19, pp. 29-41; Klapholz, B., Dietrich, B.H., Schaffner, C., CAF-1 is required for efficient replication of euchromatic DNA in Drosophila larval endocycling cells (2009) Chromosoma, 118, pp. 235-248; Kaufman, P.D., Kobayashi, R., Kessler, N., Stillman, B., The p150 and p60 subunits of chromatin assembly factor I: A molecular link between newly synthesized histones and DNA replication (1995) Cell, 81, pp. 1105-1114; Hoek, M., Stillman, B., Chromatin assembly factor 1 is essential and couples chromatin assembly to DNA replication in vivo (2003) Proceedings of the National Academy of Sciences of the United States of America, 100 (21), pp. 12183-12188. , DOI 10.1073/pnas.1635158100; Nabatiyan, A., Krude, T., Silencing of chromatin assembly factor 1 in human cells leads to cell death and loss of chromatin assembly during DNA synthesis (2004) Molecular and Cellular Biology, 24 (7), pp. 2853-2862. , DOI 10.1128/MCB.24.7.2853-2862.2004; Quivy, J.P., Gerard, A., Cook, A.J., Roche, D., Almouzni, G., The HP1-p150/CAF-1 interaction is required for pericentric heterochromatin replication and S-phase progression in mouse cells (2008) Nat Struct Mol Biol, 15, pp. 972-979; Qian, Y.-W., Wang, Y.-C.J., Hollingsworth Jr., R.E., Jones, D., Ling, N., Lee -, E.Y.H.P., A retinoblastoma-binding protein related to a negative regulator of Ras in yeast (1993) Nature, 364 (6438), pp. 648-652. , DOI 10.1038/364648a0; Parthun, M.R., Widom, J., Gottschling, D.E., The major cytoplasmic histone acetyltransferase in yeast: Links to chromatin replication and histone metabolism (1996) Cell, 87 (1), pp. 85-94. , DOI 10.1016/S0092-8674(00)81325-2; Shibahara, K.-I., Stillman, B., Replication-dependent marking of DNA by PCNA facilitates CAF-1-coupled inheritance of chromatin (1999) Cell, 96 (4), pp. 575-585; Krude, T., Chromatin assembly factor 1 (CAF-1) colocalizes with replication foci in HeLa cell nuclei (1995) Exp Cell Res, 220, pp. 304-311; Marheineke, K., Krude, T., Nucleosome assembly activity and intracellular localization of human CAF-1 changes during the cell division cycle (1998) Journal of Biological Chemistry, 273 (24), pp. 15279-15286. , DOI 10.1074/jbc.273.24.15279; Probst, A.V., Dunleavy, E., Almouzni, G., Epigenetic inheritance during the cell cycle (2009) Nat Rev Mol Cell Biol, 10, pp. 192-206; Jiao, R., Harrigan, J.A., Shevelev, I., Dietschy, T., Selak, N., Indig, F.E., Piotrowski, J., Stagljar, I., The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage (2007) Oncogene, 26 (26), pp. 3811-3822. , DOI 10.1038/sj.onc.1210150, PII 1210150; Stillman, B., Chromatin assembly during SV40 DNA replication in vitro (1986) Cell, 45 (4), pp. 555-565; Murphy, E., Shenk, T., Human cytomegalovirus genome (2008) Curr Top Microbiol Immunol, 325, pp. 1-19; Mocarski, E.S., Shenk, T., Pass, R.F., Cytomegaloviruses (2007) Fields Virology, pp. 2701-2772. , In: Knipe DM, Howley PM, eds Philadelphia: Lippincott Williams and Wilkins; Wiebusch, L., Hagemeier, C., Human cytomegalovirus 86-kilodalton IE2 protein blocks cell cycle progression in G 1 (1999) Journal of Virology, 73 (11), pp. 9274-9283; Dittmer, D., Mocarski, E.S., Human cytomegalovirus infection inhibits G 1/S transition (1997) Journal of Virology, 71 (2), pp. 1629-1634; Lu, M., Shenk, T., Human cytomegalovirus infection inhibits cell cycle progression at multiple points, including the transition from G 1 to S (1996) Journal of Virology, 70 (12), pp. 8850-8857; Estes, J.E., Huang, E.S., Stimulation of cellular thymidine kinases by human cytomegalovirus (1977) Journal of Virology, 24 (1), pp. 13-21; Masse, M.J., Karlin, S., Schachtel, G.A., Mocarski, E.S., Human cytomegalovirus origin of DNA replication (oriLyt) resides within a highly complex repetitive region (1992) Proc Natl Acad Sci USA, 89, pp. 5246-5250; Hamzeh, F.M., Lietman, P.S., Gibson, W., Hayward, G.S., Identification of the lytic origin of DNA replication in human cytomegalovirus by a novel approach utilizing ganciclovirinduced chain termination (1990) J Virol, 64, pp. 6184-6195; Borst, E.-M., Messerle, M., Analysis of human cytomegalovirus oriLyt sequence requirements in the context of the viral genome (2005) Journal of Virology, 79 (6), pp. 3615-3626. , DOI 10.1128/JVI.79.6.3615-3626.2005; Crough, T., Khanna, R., Immunobiology of human cytomegalovirus: From bench to bedside (2009) Clin Microbiol Rev, 22, pp. 76-98. , Table of Contents; Maul, G.G., Initiation of cytomegalovirus infection at ND10 (2008) Curr Top Microbiol Immunol, 325, pp. 117-132; Ahn, J.H., Jang, W.J., Hayward, G.S., The human cytomegalovirus IE2 and UL112-113 proteins accumulate in viral DNA replication compartments that initiate from the periphery of promyelocytic leukemia protein-associated nuclear bodies (PODs or ND10) (1999) J Virol, 73, pp. 10458-10471; Roizman, B., Knipe, D.M., Whitley, R.J., Hepes simplex viruses (2007) Fields Virology, pp. 2501-2601. , In: Knipe DM, Howley PM, eds Philadelphia: Lippincott Williams and Wilkins; Pari, G.S., Nuts and bolts of human cytomegalovirus lytic DNA replication (2008) Curr Top Microbiol Immunol, 325, pp. 153-166; Mercorelli, B., Sinigalia, E., Loregian, A., Palu, G., Human cytomegalovirus DNA replication: Antiviral targets and drugs (2008) Reviews in Medical Virology, 18 (3), pp. 177-210. , DOI 10.1002/rmv.558; Sourvinos, G., Tavalai, N., Berndt, A., Spandidos, D.A., Stamminger, T., Recruitment of human cytomegalovirus immediate-early 2 protein onto parental viral genomes in association with ND10 in live-infected cells (2007) Journal of Virology, 81 (18), pp. 10123-10136. , DOI 10.1128/JVI.01009-07; Sarisky, R.T., Hayward, G.S., Evidence that the UL84 gene product of human cytomegalovirus is essential for promoting oriLyt-dependent DNA replication and formation of replication compartments in cotransfection assays (1996) Journal of Virology, 70 (11), pp. 7398-7413; Marchini, A., Liu, H., Zhu, H., Human cytomegalovirus with IE-2 (UL122) deleted fails to express early lytic genes (2001) Journal of Virology, 75 (4), pp. 1870-1878. , DOI 10.1128/JVI.75.4.1870-1878.2001; Stinski, M.F., Petrik, D.T., Functional roles of the human cytomegalovirus essential IE86 protein (2008) Curr Top Microbiol Immunol, 325, pp. 133-152; Colletti, K.S., Smallenburg, K.E., Xu, Y., Pari, G.S., Human cytomegalovirus UL84 interacts with an RNA stem-loop sequence found within the RNA/DNA hybrid region of oriLyt (2007) Journal of Virology, 81 (13), pp. 7077-7085. , DOI 10.1128/JVI.00058-07; Spector, D.J., Tevethia, M.J., Protein-protein interactions between human cytomegalovirus IE2-580aa and pUL84 in lytically infected cells (1994) Journal of Virology, 68 (11), pp. 7549-7553; Xu, Y., Cei, S.A., Huete, A.R., Colletti, K.S., Pari, G.S., Human cytomegaloviras DNA replication requires transcriptional activation via an IE2- and UL84-responsive bidirectional promoter element within oriLyt (2004) Journal of Virology, 78 (21), pp. 11664-11677. , DOI 10.1128/JVI.78.21.11664-11677.2004; Petrik, D.T., Schmitt, K.P., Stinski, M.F., Inhibition of cellular DNA synthesis by the human cytomegalovirus IE86 protein is necessary for efficient virus replication (2006) J Virol, 80, pp. 3872-3883; Kent, J.R., Zeng, P.-Y., Atanasiu, D., Gardner, J., Fraser, N.W., Berger, S.L., During lytic infection herpes simplex virus type 1 is associated with histones bearing modifications that correlate with active transcription (2004) Journal of Virology, 78 (18), pp. 10178-10186. , DOI 10.1128/JVI.78.18.10178-10186.2004; Deshmane, S.L., Fraser, N.W., During latency, herpes simplex virus type 1 DNA is associated with nucleosomes in a chromatin structure (1989) Journal of Virology, 63 (2), pp. 943-947; Nitzsche, A., Paulus, C., Nevels, M., Temporal dynamics of cytomegalovirus chromatin assembly in productively infected human cells (2008) J Virol, 82, pp. 11167-11180; Mate, J.L., Ariza, A., Munoz, A., Molinero, J.L., Lopez, D., Navas-Palacios, J.J., Induction of proliferating cell nuclear antigen and Ki-67 expression by cytomegalovirus infection (1998) Journal of Pathology, 184 (3), pp. 279-282. , DOI 10.1002/(SICI)1096-9896(199803)184:3<279::AID-PATH7>3.0.CO;2-4; Huang, C.F., Wang, Y.C., Tsao, D.A., Tung, S.F., Lin, Y.S., Wu, C.W., Antagonism between members of the CNC-bZIP family and the immediate-early protein IE2 of human cytomegalovirus (2000) J Biol Chem, 275, pp. 12313-12320; Nakayasu, H., Berezney, R., Mapping replicational sites in the eucaryotic cell nucleus (1989) J Cell Biol, 108, pp. 1-11; Van Dierendonck, J.H., Keyzer, R., Van De Velde, C.J.H., Cornelisse, C.J., Subdivision of S-phase by analysis of nuclear 5-bromodeoxyuridine staining patterns (1989) Cytometry, 10 (2), pp. 143-150. , DOI 10.1002/cyto.990100205; Castillo, J.P., Kowalik, T.F., Human cytomegalovirus immediate early proteins and cell growth control (2002) Gene, 290 (1-2), pp. 19-34. , DOI 10.1016/S0378-1119(02)00566-8, PII S0378111902005668; Reeves, M., Murphy, J., Greaves, R., Fairley, J., Brehm, A., Sinclair, J., Autorepression of the human cytomegalovirus major immediate-early promoter/enhancer at late times of infection is mediated by the recruitment of chromatin remodeling enzymes by IE86 (2006) Journal of Virology, 80 (20), pp. 9998-10009. , DOI 10.1128/JVI.01297-06; Huang, E.S., Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid (1975) J Virol, 16, pp. 1560-1565; Ye, X., Franco, A.A., Santos, H., Nelson, D.M., Kaufman, P.D., Adams, P.D., Defective S phase chromatin assembly causes DNA damage, activation of the S phase checkpoint, and S phase arrest (2003) Molecular Cell, 11 (2), pp. 341-351. , DOI 10.1016/S1097-2765(03)00037-6; Colletti, K.S., Xu, Y., Cei, S.A., Tarrant, M., Pari, G.S., Human cytomegalovirus UL84 oligomerization and heterodimerization domains act as transdominant inhibitors of oriLyt-dependent DNA replication: Evidence that IE2-UL84 and UL84-UL84 interactions are required for lytic DNA replication (2004) Journal of Virology, 78 (17), pp. 9203-9214. , DOI 10.1128/JVI.78.17.9203-9214.2004; Loyola, A., Bonaldi, T., Roche, D., Imhof, A., Almouzni, G., PTMs on H3 variants before chromatin assembly potentiate their final epigenetic state (2006) Molecular Cell, 24 (2), pp. 309-316. , DOI 10.1016/j.molcel.2006.08.019, PII S1097276506006009; Lee, S.B., Ou, D.S., Lee, C.F., Juan, L.J., Gene-specific transcriptional activation mediated by the p150 subunit of the chromatin assembly factor 1 (2009) J Biol Chem, 284, pp. 14040-14049; Baldick Jr., C.J., Shenk, T., Proteins associated with purified human cytomegalovirus particles (1996) Journal of Virology, 70 (9), pp. 6097-6105; Varnum, S.M., Streblow, D.N., Monroe, M.E., Smith, P., Auberry, K.J., Pasa-Tolic, L., Wang, D., Nelson, J.A., Identification of proteins in human cytomegalovirus (HCMV) particles: The HCMV proteome (2004) Journal of Virology, 78 (20), pp. 10960-10966. , DOI 10.1128/JVI.78.20.10960-10966.2004; Groves, I., Reeves, M., Sinclair, J., Lytic infection of permissive cells with human cytomegalovirus is regulated by an intrinsic {"}pre-immediate early{"} repression of viral gene expression mediated by histone post-translational modification (2009) J Gen Virol, 90, pp. 2364-2374; Liu, X.-F., Yan, S., Abecassis, M., Hummel, M., Establishment of murine cytomegalovirus latency in vivo is associated with changes in histone modifications and recruitment of transcriptional repressors to the major immediate-early promoter (2008) Journal of Virology, 82 (21), pp. 10922-10931. , http://jvi.asm.org/cgi/reprint/82/21/10922, DOI 10.1128/JVI.00865-08; Cuevas-Bennett, C., Shenk, T., Dynamic histone H3 acetylation and methylation at human cytomegalovirus promoters during replication in fibroblasts (2008) J Virol, 82, pp. 9525-9536; Ioudinkova, E., Arcangeletti, M.C., Rynditch, A., De Conto, F., Motta, F., Covan, S., Pinardi, F., Chezzi, C., Control of human cytomegalovirus gene expression by differential histone modifications during lytic and latent infection of a monocytic cell line (2006) Gene, 384 (1-2), pp. 120-128. , DOI 10.1016/j.gene.2006.07.021, PII S0378111906004719; Reeves, M.B., MacAry, P.A., Lehner, P.J., Sissons, J.G.P., Sinclair, J.H., Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers (2005) Proceedings of the National Academy of Sciences of the United States of America, 102 (11), pp. 4140-4145. , DOI 10.1073/pnas.0408994102; Murphy, J.C., Fischle, W., Verdin, E., Sinclair, J.H., Control of cytomegalovirus lytic gene expression by histone acetylation (2002) EMBO Journal, 21 (5), pp. 1112-1120. , DOI 10.1093/emboj/21.5.1112; Knipe, D.M., Cliffe, A., Chromatin control of herpes simplex virus lytic and latent infection (2008) Nature Reviews Microbiology, 6 (3), pp. 211-221. , DOI 10.1038/nrmicro1794, PII NRMICRO1794; Peng, H., Nogueira, M.L., Vogel, J.L., Kristie, T.M., Transcriptional coactivator HCF-1 couples the histone chaperone Asf1b to HSV-1 DNA replication components Proc Natl Acad Sci USA, 107, pp. 2461-2466; Sanders, R.L., Del Rosario, C.J., White, E.A., Spector, D.H., Internal deletions of IE2 86 and loss of the late IE2 60 and IE2 40 proteins encoded by human cytomegalovirus affect the levels of UL84 protein but not the amount of UL84 mRNA or the loading and distribution of the mRNA on polysomes (2008) J Virol, 82, pp. 11383-11397; Kierszenbaum, A.L., Huang, E.S., Chromatin pattern consisting of repeating bipartite structures in WI-38 cells infected with human cytomegalovirus (1978) Journal of Virology, 28 (2), pp. 661-664; Salvant, B.S., Fortunato, E.A., Spector, D.H., Cell cycle dysregulation by human cytomegalovirus: Influence of the cell cycle phase at the time of infection and effects on cyclin transcription (1998) Journal of Virology, 72 (5), pp. 3729-3741; Bresnahan, W.A., Boldogh, I., Thompson, E.A., Albrecht, T., Human cytomegalovirus inhibits cellular DNA synthesis and arrests productively infected cells in late G1 (1996) Virology, 224 (1), pp. 150-160. , DOI 10.1006/viro.1996.0516; Jault, F.M., Jault, J.M., Ruchti, F., Cytomegalovirus infection induces high levels of cyclins, phosphorylated Rb, and p53, leading to cell cycle arrest (1995) J Virol, 69, pp. 6697-6704; Huang, S., Zhou, H., Tarara, J., Zhang, Z., A novel role for histone chaperones CAF-1 and Rtt106p in heterochromatin silencing (2007) EMBO Journal, 26 (9), pp. 2274-2283. , DOI 10.1038/sj.emboj.7601670, PII 7601670; De Koning, L., Corpet, A., Haber, J.E., Almouzni, G., Histone chaperones: An escort network regulating histone traffic (2007) Nature Structural and Molecular Biology, 14 (11), pp. 997-1007. , DOI 10.1038/nsmb1318, PII NSMB1318; Spaete, R.R., Mocarski, E.S., Insertion and deletion mutagenesis of the human cytomegalovirus genome (1987) Proc Natl Acad Sci USA, 84, pp. 7213-7217; Lee, C.F., Ou, D.S., Lee, S.B., HNaa10p contributes to tumorigenesis by facilitating DNMT1-mediated tumor suppressor gene silencing (2010) J Clin Invest, 120, pp. 2920-2930",
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TY - JOUR

T1 - Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2

AU - Lee, Sung-Bau

AU - Lee, Chung-Fan

AU - Ou, Derick S C

AU - Dulal, Kalpana

AU - Chang, Liang-Hao

AU - Ma, Chen-Han

AU - Huang, Chien-Fu

AU - Zhu, Hua

AU - Lin, Young-Sun

AU - Juan, Li-Jung

N1 - 被引用次數:4 Export Date: 28 March 2016 通訊地址: Juan, L.-J.; Genomics Research Center, Academia Sinica, Taipei 115, Taiwan; 電子郵件: ljjuan@gate.sinica.edu.tw 化學物質/CAS: histone, 9062-68-4; Chromatin; Chromatin Assembly Factor-1; DNA, Viral; Histones; IE2 protein, Cytomegalovirus; Immediate-Early Proteins; Protein Subunits; Trans-Activators 參考文獻: Smith, S., Stillman, B., Purification and characterization of CAF-I, a human cell factor required for chromatin assembly during DNA replication in vitro (1989) Cell, 58 (1), pp. 15-25. , DOI 10.1016/0092-8674(89)90398-X; Li, Q., Zhou, H., Wurtele, H., Davies, B., Horazdovsky, B., Verreault, A., Zhang, Z., Acetylation of histone H3 lysine 56 regulates replication-coupled nucleosome assembly (2008) Cell, 134 (2), pp. 244-255. , DOI 10.1016/j.cell.2008.06.018, PII S0092867408007708; Takami, Y., Ono, T., Fukagawa, T., Shibahara, K.-I., Nakayama, T., Essential role of chromatin assembly factor-1-mediated rapid nucleosome assembly for DNA replication and cell division in vertebrate cells (2007) Molecular Biology of the Cell, 18 (1), pp. 129-141. , DOI 10.1091/mbc.E06-05-0426; Corpet, A., Almouzni, G., Making copies of chromatin: The challenge of nucleosomal organization and epigenetic information (2009) Trends Cell Biol, 19, pp. 29-41; Klapholz, B., Dietrich, B.H., Schaffner, C., CAF-1 is required for efficient replication of euchromatic DNA in Drosophila larval endocycling cells (2009) Chromosoma, 118, pp. 235-248; Kaufman, P.D., Kobayashi, R., Kessler, N., Stillman, B., The p150 and p60 subunits of chromatin assembly factor I: A molecular link between newly synthesized histones and DNA replication (1995) Cell, 81, pp. 1105-1114; Hoek, M., Stillman, B., Chromatin assembly factor 1 is essential and couples chromatin assembly to DNA replication in vivo (2003) Proceedings of the National Academy of Sciences of the United States of America, 100 (21), pp. 12183-12188. , DOI 10.1073/pnas.1635158100; Nabatiyan, A., Krude, T., Silencing of chromatin assembly factor 1 in human cells leads to cell death and loss of chromatin assembly during DNA synthesis (2004) Molecular and Cellular Biology, 24 (7), pp. 2853-2862. , DOI 10.1128/MCB.24.7.2853-2862.2004; Quivy, J.P., Gerard, A., Cook, A.J., Roche, D., Almouzni, G., The HP1-p150/CAF-1 interaction is required for pericentric heterochromatin replication and S-phase progression in mouse cells (2008) Nat Struct Mol Biol, 15, pp. 972-979; Qian, Y.-W., Wang, Y.-C.J., Hollingsworth Jr., R.E., Jones, D., Ling, N., Lee -, E.Y.H.P., A retinoblastoma-binding protein related to a negative regulator of Ras in yeast (1993) Nature, 364 (6438), pp. 648-652. , DOI 10.1038/364648a0; Parthun, M.R., Widom, J., Gottschling, D.E., The major cytoplasmic histone acetyltransferase in yeast: Links to chromatin replication and histone metabolism (1996) Cell, 87 (1), pp. 85-94. , DOI 10.1016/S0092-8674(00)81325-2; Shibahara, K.-I., Stillman, B., Replication-dependent marking of DNA by PCNA facilitates CAF-1-coupled inheritance of chromatin (1999) Cell, 96 (4), pp. 575-585; Krude, T., Chromatin assembly factor 1 (CAF-1) colocalizes with replication foci in HeLa cell nuclei (1995) Exp Cell Res, 220, pp. 304-311; Marheineke, K., Krude, T., Nucleosome assembly activity and intracellular localization of human CAF-1 changes during the cell division cycle (1998) Journal of Biological Chemistry, 273 (24), pp. 15279-15286. , DOI 10.1074/jbc.273.24.15279; Probst, A.V., Dunleavy, E., Almouzni, G., Epigenetic inheritance during the cell cycle (2009) Nat Rev Mol Cell Biol, 10, pp. 192-206; Jiao, R., Harrigan, J.A., Shevelev, I., Dietschy, T., Selak, N., Indig, F.E., Piotrowski, J., Stagljar, I., The Werner syndrome protein is required for recruitment of chromatin assembly factor 1 following DNA damage (2007) Oncogene, 26 (26), pp. 3811-3822. , DOI 10.1038/sj.onc.1210150, PII 1210150; Stillman, B., Chromatin assembly during SV40 DNA replication in vitro (1986) Cell, 45 (4), pp. 555-565; Murphy, E., Shenk, T., Human cytomegalovirus genome (2008) Curr Top Microbiol Immunol, 325, pp. 1-19; Mocarski, E.S., Shenk, T., Pass, R.F., Cytomegaloviruses (2007) Fields Virology, pp. 2701-2772. , In: Knipe DM, Howley PM, eds Philadelphia: Lippincott Williams and Wilkins; Wiebusch, L., Hagemeier, C., Human cytomegalovirus 86-kilodalton IE2 protein blocks cell cycle progression in G 1 (1999) Journal of Virology, 73 (11), pp. 9274-9283; Dittmer, D., Mocarski, E.S., Human cytomegalovirus infection inhibits G 1/S transition (1997) Journal of Virology, 71 (2), pp. 1629-1634; Lu, M., Shenk, T., Human cytomegalovirus infection inhibits cell cycle progression at multiple points, including the transition from G 1 to S (1996) Journal of Virology, 70 (12), pp. 8850-8857; Estes, J.E., Huang, E.S., Stimulation of cellular thymidine kinases by human cytomegalovirus (1977) Journal of Virology, 24 (1), pp. 13-21; Masse, M.J., Karlin, S., Schachtel, G.A., Mocarski, E.S., Human cytomegalovirus origin of DNA replication (oriLyt) resides within a highly complex repetitive region (1992) Proc Natl Acad Sci USA, 89, pp. 5246-5250; Hamzeh, F.M., Lietman, P.S., Gibson, W., Hayward, G.S., Identification of the lytic origin of DNA replication in human cytomegalovirus by a novel approach utilizing ganciclovirinduced chain termination (1990) J Virol, 64, pp. 6184-6195; Borst, E.-M., Messerle, M., Analysis of human cytomegalovirus oriLyt sequence requirements in the context of the viral genome (2005) Journal of Virology, 79 (6), pp. 3615-3626. , DOI 10.1128/JVI.79.6.3615-3626.2005; Crough, T., Khanna, R., Immunobiology of human cytomegalovirus: From bench to bedside (2009) Clin Microbiol Rev, 22, pp. 76-98. , Table of Contents; Maul, G.G., Initiation of cytomegalovirus infection at ND10 (2008) Curr Top Microbiol Immunol, 325, pp. 117-132; Ahn, J.H., Jang, W.J., Hayward, G.S., The human cytomegalovirus IE2 and UL112-113 proteins accumulate in viral DNA replication compartments that initiate from the periphery of promyelocytic leukemia protein-associated nuclear bodies (PODs or ND10) (1999) J Virol, 73, pp. 10458-10471; Roizman, B., Knipe, D.M., Whitley, R.J., Hepes simplex viruses (2007) Fields Virology, pp. 2501-2601. , In: Knipe DM, Howley PM, eds Philadelphia: Lippincott Williams and Wilkins; Pari, G.S., Nuts and bolts of human cytomegalovirus lytic DNA replication (2008) Curr Top Microbiol Immunol, 325, pp. 153-166; Mercorelli, B., Sinigalia, E., Loregian, A., Palu, G., Human cytomegalovirus DNA replication: Antiviral targets and drugs (2008) Reviews in Medical Virology, 18 (3), pp. 177-210. , DOI 10.1002/rmv.558; Sourvinos, G., Tavalai, N., Berndt, A., Spandidos, D.A., Stamminger, T., Recruitment of human cytomegalovirus immediate-early 2 protein onto parental viral genomes in association with ND10 in live-infected cells (2007) Journal of Virology, 81 (18), pp. 10123-10136. , DOI 10.1128/JVI.01009-07; Sarisky, R.T., Hayward, G.S., Evidence that the UL84 gene product of human cytomegalovirus is essential for promoting oriLyt-dependent DNA replication and formation of replication compartments in cotransfection assays (1996) Journal of Virology, 70 (11), pp. 7398-7413; Marchini, A., Liu, H., Zhu, H., Human cytomegalovirus with IE-2 (UL122) deleted fails to express early lytic genes (2001) Journal of Virology, 75 (4), pp. 1870-1878. , DOI 10.1128/JVI.75.4.1870-1878.2001; Stinski, M.F., Petrik, D.T., Functional roles of the human cytomegalovirus essential IE86 protein (2008) Curr Top Microbiol Immunol, 325, pp. 133-152; Colletti, K.S., Smallenburg, K.E., Xu, Y., Pari, G.S., Human cytomegalovirus UL84 interacts with an RNA stem-loop sequence found within the RNA/DNA hybrid region of oriLyt (2007) Journal of Virology, 81 (13), pp. 7077-7085. , DOI 10.1128/JVI.00058-07; Spector, D.J., Tevethia, M.J., Protein-protein interactions between human cytomegalovirus IE2-580aa and pUL84 in lytically infected cells (1994) Journal of Virology, 68 (11), pp. 7549-7553; Xu, Y., Cei, S.A., Huete, A.R., Colletti, K.S., Pari, G.S., Human cytomegaloviras DNA replication requires transcriptional activation via an IE2- and UL84-responsive bidirectional promoter element within oriLyt (2004) Journal of Virology, 78 (21), pp. 11664-11677. , DOI 10.1128/JVI.78.21.11664-11677.2004; Petrik, D.T., Schmitt, K.P., Stinski, M.F., Inhibition of cellular DNA synthesis by the human cytomegalovirus IE86 protein is necessary for efficient virus replication (2006) J Virol, 80, pp. 3872-3883; Kent, J.R., Zeng, P.-Y., Atanasiu, D., Gardner, J., Fraser, N.W., Berger, S.L., During lytic infection herpes simplex virus type 1 is associated with histones bearing modifications that correlate with active transcription (2004) Journal of Virology, 78 (18), pp. 10178-10186. , DOI 10.1128/JVI.78.18.10178-10186.2004; Deshmane, S.L., Fraser, N.W., During latency, herpes simplex virus type 1 DNA is associated with nucleosomes in a chromatin structure (1989) Journal of Virology, 63 (2), pp. 943-947; Nitzsche, A., Paulus, C., Nevels, M., Temporal dynamics of cytomegalovirus chromatin assembly in productively infected human cells (2008) J Virol, 82, pp. 11167-11180; Mate, J.L., Ariza, A., Munoz, A., Molinero, J.L., Lopez, D., Navas-Palacios, J.J., Induction of proliferating cell nuclear antigen and Ki-67 expression by cytomegalovirus infection (1998) Journal of Pathology, 184 (3), pp. 279-282. , DOI 10.1002/(SICI)1096-9896(199803)184:3<279::AID-PATH7>3.0.CO;2-4; Huang, C.F., Wang, Y.C., Tsao, D.A., Tung, S.F., Lin, Y.S., Wu, C.W., Antagonism between members of the CNC-bZIP family and the immediate-early protein IE2 of human cytomegalovirus (2000) J Biol Chem, 275, pp. 12313-12320; Nakayasu, H., Berezney, R., Mapping replicational sites in the eucaryotic cell nucleus (1989) J Cell Biol, 108, pp. 1-11; Van Dierendonck, J.H., Keyzer, R., Van De Velde, C.J.H., Cornelisse, C.J., Subdivision of S-phase by analysis of nuclear 5-bromodeoxyuridine staining patterns (1989) Cytometry, 10 (2), pp. 143-150. , DOI 10.1002/cyto.990100205; Castillo, J.P., Kowalik, T.F., Human cytomegalovirus immediate early proteins and cell growth control (2002) Gene, 290 (1-2), pp. 19-34. , DOI 10.1016/S0378-1119(02)00566-8, PII S0378111902005668; Reeves, M., Murphy, J., Greaves, R., Fairley, J., Brehm, A., Sinclair, J., Autorepression of the human cytomegalovirus major immediate-early promoter/enhancer at late times of infection is mediated by the recruitment of chromatin remodeling enzymes by IE86 (2006) Journal of Virology, 80 (20), pp. 9998-10009. , DOI 10.1128/JVI.01297-06; Huang, E.S., Human cytomegalovirus. IV. Specific inhibition of virus-induced DNA polymerase activity and viral DNA replication by phosphonoacetic acid (1975) J Virol, 16, pp. 1560-1565; Ye, X., Franco, A.A., Santos, H., Nelson, D.M., Kaufman, P.D., Adams, P.D., Defective S phase chromatin assembly causes DNA damage, activation of the S phase checkpoint, and S phase arrest (2003) Molecular Cell, 11 (2), pp. 341-351. , DOI 10.1016/S1097-2765(03)00037-6; Colletti, K.S., Xu, Y., Cei, S.A., Tarrant, M., Pari, G.S., Human cytomegalovirus UL84 oligomerization and heterodimerization domains act as transdominant inhibitors of oriLyt-dependent DNA replication: Evidence that IE2-UL84 and UL84-UL84 interactions are required for lytic DNA replication (2004) Journal of Virology, 78 (17), pp. 9203-9214. , DOI 10.1128/JVI.78.17.9203-9214.2004; Loyola, A., Bonaldi, T., Roche, D., Imhof, A., Almouzni, G., PTMs on H3 variants before chromatin assembly potentiate their final epigenetic state (2006) Molecular Cell, 24 (2), pp. 309-316. , DOI 10.1016/j.molcel.2006.08.019, PII S1097276506006009; Lee, S.B., Ou, D.S., Lee, C.F., Juan, L.J., Gene-specific transcriptional activation mediated by the p150 subunit of the chromatin assembly factor 1 (2009) J Biol Chem, 284, pp. 14040-14049; Baldick Jr., C.J., Shenk, T., Proteins associated with purified human cytomegalovirus particles (1996) Journal of Virology, 70 (9), pp. 6097-6105; Varnum, S.M., Streblow, D.N., Monroe, M.E., Smith, P., Auberry, K.J., Pasa-Tolic, L., Wang, D., Nelson, J.A., Identification of proteins in human cytomegalovirus (HCMV) particles: The HCMV proteome (2004) Journal of Virology, 78 (20), pp. 10960-10966. , DOI 10.1128/JVI.78.20.10960-10966.2004; Groves, I., Reeves, M., Sinclair, J., Lytic infection of permissive cells with human cytomegalovirus is regulated by an intrinsic "pre-immediate early" repression of viral gene expression mediated by histone post-translational modification (2009) J Gen Virol, 90, pp. 2364-2374; Liu, X.-F., Yan, S., Abecassis, M., Hummel, M., Establishment of murine cytomegalovirus latency in vivo is associated with changes in histone modifications and recruitment of transcriptional repressors to the major immediate-early promoter (2008) Journal of Virology, 82 (21), pp. 10922-10931. , http://jvi.asm.org/cgi/reprint/82/21/10922, DOI 10.1128/JVI.00865-08; Cuevas-Bennett, C., Shenk, T., Dynamic histone H3 acetylation and methylation at human cytomegalovirus promoters during replication in fibroblasts (2008) J Virol, 82, pp. 9525-9536; Ioudinkova, E., Arcangeletti, M.C., Rynditch, A., De Conto, F., Motta, F., Covan, S., Pinardi, F., Chezzi, C., Control of human cytomegalovirus gene expression by differential histone modifications during lytic and latent infection of a monocytic cell line (2006) Gene, 384 (1-2), pp. 120-128. , DOI 10.1016/j.gene.2006.07.021, PII S0378111906004719; Reeves, M.B., MacAry, P.A., Lehner, P.J., Sissons, J.G.P., Sinclair, J.H., Latency, chromatin remodeling, and reactivation of human cytomegalovirus in the dendritic cells of healthy carriers (2005) Proceedings of the National Academy of Sciences of the United States of America, 102 (11), pp. 4140-4145. , DOI 10.1073/pnas.0408994102; Murphy, J.C., Fischle, W., Verdin, E., Sinclair, J.H., Control of cytomegalovirus lytic gene expression by histone acetylation (2002) EMBO Journal, 21 (5), pp. 1112-1120. , DOI 10.1093/emboj/21.5.1112; Knipe, D.M., Cliffe, A., Chromatin control of herpes simplex virus lytic and latent infection (2008) Nature Reviews Microbiology, 6 (3), pp. 211-221. , DOI 10.1038/nrmicro1794, PII NRMICRO1794; Peng, H., Nogueira, M.L., Vogel, J.L., Kristie, T.M., Transcriptional coactivator HCF-1 couples the histone chaperone Asf1b to HSV-1 DNA replication components Proc Natl Acad Sci USA, 107, pp. 2461-2466; Sanders, R.L., Del Rosario, C.J., White, E.A., Spector, D.H., Internal deletions of IE2 86 and loss of the late IE2 60 and IE2 40 proteins encoded by human cytomegalovirus affect the levels of UL84 protein but not the amount of UL84 mRNA or the loading and distribution of the mRNA on polysomes (2008) J Virol, 82, pp. 11383-11397; Kierszenbaum, A.L., Huang, E.S., Chromatin pattern consisting of repeating bipartite structures in WI-38 cells infected with human cytomegalovirus (1978) Journal of Virology, 28 (2), pp. 661-664; Salvant, B.S., Fortunato, E.A., Spector, D.H., Cell cycle dysregulation by human cytomegalovirus: Influence of the cell cycle phase at the time of infection and effects on cyclin transcription (1998) Journal of Virology, 72 (5), pp. 3729-3741; Bresnahan, W.A., Boldogh, I., Thompson, E.A., Albrecht, T., Human cytomegalovirus inhibits cellular DNA synthesis and arrests productively infected cells in late G1 (1996) Virology, 224 (1), pp. 150-160. , DOI 10.1006/viro.1996.0516; Jault, F.M., Jault, J.M., Ruchti, F., Cytomegalovirus infection induces high levels of cyclins, phosphorylated Rb, and p53, leading to cell cycle arrest (1995) J Virol, 69, pp. 6697-6704; Huang, S., Zhou, H., Tarara, J., Zhang, Z., A novel role for histone chaperones CAF-1 and Rtt106p in heterochromatin silencing (2007) EMBO Journal, 26 (9), pp. 2274-2283. , DOI 10.1038/sj.emboj.7601670, PII 7601670; De Koning, L., Corpet, A., Haber, J.E., Almouzni, G., Histone chaperones: An escort network regulating histone traffic (2007) Nature Structural and Molecular Biology, 14 (11), pp. 997-1007. , DOI 10.1038/nsmb1318, PII NSMB1318; Spaete, R.R., Mocarski, E.S., Insertion and deletion mutagenesis of the human cytomegalovirus genome (1987) Proc Natl Acad Sci USA, 84, pp. 7213-7217; Lee, C.F., Ou, D.S., Lee, S.B., HNaa10p contributes to tumorigenesis by facilitating DNMT1-mediated tumor suppressor gene silencing (2010) J Clin Invest, 120, pp. 2920-2930

PY - 2011

Y1 - 2011

N2 - Chromatin assembly factor 1 (CAF1) consisting of p150, p60 and p48 is known to assemble histones onto newly synthesized DNA and thus maintain the chromatin structure. Here, we show that CAF1 expression was induced in human cytomegalovirus (HCMV)-infected cells, concomitantly with global chromatin decondensation. This apparent conflict was thought to result, in part, from CAF1 mislocalization to compartments of HCMV DNA synthesis through binding of its largest subunit p150 to viral immediate-early protein 2 (IE2). p150 interaction with p60 and IE2 facilitated HCMV DNA synthesis. The IE2Q548R mutation, previously reported to result in impaired HCMV growth with unknown mechanism, disrupted IE2/p150 and IE2/histones association in our study. Moreover, IE2 interaction with histones partly depends on p150, and the HCMV-induced chromatin decondensation was reduced in cells ectopically expressing the p150 mutant defective in IE2 binding. These results not only indicate that CAF1 was hijacked by IE2 to facilitate the replication of the HCMV genome, suggesting chromatin assembly plays an important role in herpesviral DNA synthesis, but also provide a model of the virus-induced chromatin instability through CAF1. © 2011 IBCB, SIBS, CAS All rights reserved.

AB - Chromatin assembly factor 1 (CAF1) consisting of p150, p60 and p48 is known to assemble histones onto newly synthesized DNA and thus maintain the chromatin structure. Here, we show that CAF1 expression was induced in human cytomegalovirus (HCMV)-infected cells, concomitantly with global chromatin decondensation. This apparent conflict was thought to result, in part, from CAF1 mislocalization to compartments of HCMV DNA synthesis through binding of its largest subunit p150 to viral immediate-early protein 2 (IE2). p150 interaction with p60 and IE2 facilitated HCMV DNA synthesis. The IE2Q548R mutation, previously reported to result in impaired HCMV growth with unknown mechanism, disrupted IE2/p150 and IE2/histones association in our study. Moreover, IE2 interaction with histones partly depends on p150, and the HCMV-induced chromatin decondensation was reduced in cells ectopically expressing the p150 mutant defective in IE2 binding. These results not only indicate that CAF1 was hijacked by IE2 to facilitate the replication of the HCMV genome, suggesting chromatin assembly plays an important role in herpesviral DNA synthesis, but also provide a model of the virus-induced chromatin instability through CAF1. © 2011 IBCB, SIBS, CAS All rights reserved.

KW - chromatin assembly factor 1

KW - chromatin structure

KW - human cytomegalovirus

KW - immediate-early protein 2

KW - histone

KW - IE2 protein, Cytomegalovirus

KW - immediate early protein

KW - transactivator protein

KW - virus DNA

KW - amino acid substitution

KW - article

KW - cell line

KW - chromatin

KW - chromatin assembly and disassembly

KW - Cytomegalovirus

KW - genetics

KW - host pathogen interaction

KW - human

KW - metabolism

KW - protein binding

KW - protein subunit

KW - virus replication

KW - Amino Acid Substitution

KW - Cell Line

KW - Chromatin

KW - Chromatin Assembly and Disassembly

KW - Chromatin Assembly Factor-1

KW - DNA, Viral

KW - Histones

KW - Host-Pathogen Interactions

KW - Humans

KW - Immediate-Early Proteins

KW - Protein Binding

KW - Protein Subunits

KW - Trans-Activators

KW - Virus Replication

KW - Human herpesvirus 5

U2 - 10.1038/cr.2011.53

DO - 10.1038/cr.2011.53

M3 - Article

VL - 21

SP - 1230

EP - 1247

JO - Cell Research

JF - Cell Research

SN - 1001-0602

IS - 8

ER -