Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen

Mel Campbell, Pei Ching Chang, Steve Huerta, Chie Izumiya, Ryan Davis, Clifford G. Tepper, Kevin Y. Kim, Bogdan Shevchenko, Don Hong Wang, Jae U. Jung, Paul A. Luciw, Hsing Jien Kung, Yoshihiro Izumiya

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The Kaposi sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a multifunctional protein with roles in gene regulation and maintenance of viral latency. Post-translational modification of LANA is important for functional diversification. Here, we report that LANA is subject to arginine methylation by protein arginine methyltransferase 1 in vitro and in vivo. The major arginine methylation site in LANA was mapped to arginine 20. This site was mutated to either phenylalanine (bulky hydrophobic, constitutive methylated mimetic) or lysine (positively charged, non-arginine methylatable) residues. The significance of the methylation in LANA function was examined in both the isolated form and in the context of the viral genome through the generation of recombinant KSHV. In addition, authentic LANA binding sites on the KSHV episome in naturally infected cells were identified using a whole genome KSHV tiling array. Although mutation of the methylation site resulted in no significant difference in KSHV LANA subcellular localization, we found that the methylation mimetic mutation resulted in augmented histone binding in vitro and increased LANA occupancy at identified LANA target promoters in vivo. Moreover, a cell line carrying the methylation mimetic mutant KSHV showed reduced viral gene expression relative to controls both in latency and in the course of reactivation. These results suggest that residue 20 is important for modulation of a subset of LANA functions and properties of this residue, including the hydrophobic character induced by arginine methylation, may contribute to the observed effects.

Original languageEnglish
Pages (from-to)5806-5818
Number of pages13
JournalJournal of Biological Chemistry
Volume287
Issue number8
DOIs
Publication statusPublished - Feb 17 2012
Externally publishedYes

Fingerprint

Protein-Arginine N-Methyltransferases
Human Herpesvirus 8
Methylation
Arginine
Gene expression
Genes
latency-associated nuclear antigen
Virus Latency
Mutation
Viral Genes
Viral Genome
Post Translational Protein Processing
Set theory
Phenylalanine
Histones
Lysine
Plasmids

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen. / Campbell, Mel; Chang, Pei Ching; Huerta, Steve; Izumiya, Chie; Davis, Ryan; Tepper, Clifford G.; Kim, Kevin Y.; Shevchenko, Bogdan; Wang, Don Hong; Jung, Jae U.; Luciw, Paul A.; Kung, Hsing Jien; Izumiya, Yoshihiro.

In: Journal of Biological Chemistry, Vol. 287, No. 8, 17.02.2012, p. 5806-5818.

Research output: Contribution to journalArticle

Campbell, M, Chang, PC, Huerta, S, Izumiya, C, Davis, R, Tepper, CG, Kim, KY, Shevchenko, B, Wang, DH, Jung, JU, Luciw, PA, Kung, HJ & Izumiya, Y 2012, 'Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen', Journal of Biological Chemistry, vol. 287, no. 8, pp. 5806-5818. https://doi.org/10.1074/jbc.M111.289496
Campbell, Mel ; Chang, Pei Ching ; Huerta, Steve ; Izumiya, Chie ; Davis, Ryan ; Tepper, Clifford G. ; Kim, Kevin Y. ; Shevchenko, Bogdan ; Wang, Don Hong ; Jung, Jae U. ; Luciw, Paul A. ; Kung, Hsing Jien ; Izumiya, Yoshihiro. / Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 8. pp. 5806-5818.
@article{ce9f1823192f431a93ae335a567051ce,
title = "Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen",
abstract = "The Kaposi sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a multifunctional protein with roles in gene regulation and maintenance of viral latency. Post-translational modification of LANA is important for functional diversification. Here, we report that LANA is subject to arginine methylation by protein arginine methyltransferase 1 in vitro and in vivo. The major arginine methylation site in LANA was mapped to arginine 20. This site was mutated to either phenylalanine (bulky hydrophobic, constitutive methylated mimetic) or lysine (positively charged, non-arginine methylatable) residues. The significance of the methylation in LANA function was examined in both the isolated form and in the context of the viral genome through the generation of recombinant KSHV. In addition, authentic LANA binding sites on the KSHV episome in naturally infected cells were identified using a whole genome KSHV tiling array. Although mutation of the methylation site resulted in no significant difference in KSHV LANA subcellular localization, we found that the methylation mimetic mutation resulted in augmented histone binding in vitro and increased LANA occupancy at identified LANA target promoters in vivo. Moreover, a cell line carrying the methylation mimetic mutant KSHV showed reduced viral gene expression relative to controls both in latency and in the course of reactivation. These results suggest that residue 20 is important for modulation of a subset of LANA functions and properties of this residue, including the hydrophobic character induced by arginine methylation, may contribute to the observed effects.",
author = "Mel Campbell and Chang, {Pei Ching} and Steve Huerta and Chie Izumiya and Ryan Davis and Tepper, {Clifford G.} and Kim, {Kevin Y.} and Bogdan Shevchenko and Wang, {Don Hong} and Jung, {Jae U.} and Luciw, {Paul A.} and Kung, {Hsing Jien} and Yoshihiro Izumiya",
year = "2012",
month = "2",
day = "17",
doi = "10.1074/jbc.M111.289496",
language = "English",
volume = "287",
pages = "5806--5818",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "8",

}

TY - JOUR

T1 - Protein arginine methyltransferase 1-directed methylation of Kaposi sarcoma-associated herpesvirus latency-associated nuclear antigen

AU - Campbell, Mel

AU - Chang, Pei Ching

AU - Huerta, Steve

AU - Izumiya, Chie

AU - Davis, Ryan

AU - Tepper, Clifford G.

AU - Kim, Kevin Y.

AU - Shevchenko, Bogdan

AU - Wang, Don Hong

AU - Jung, Jae U.

AU - Luciw, Paul A.

AU - Kung, Hsing Jien

AU - Izumiya, Yoshihiro

PY - 2012/2/17

Y1 - 2012/2/17

N2 - The Kaposi sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a multifunctional protein with roles in gene regulation and maintenance of viral latency. Post-translational modification of LANA is important for functional diversification. Here, we report that LANA is subject to arginine methylation by protein arginine methyltransferase 1 in vitro and in vivo. The major arginine methylation site in LANA was mapped to arginine 20. This site was mutated to either phenylalanine (bulky hydrophobic, constitutive methylated mimetic) or lysine (positively charged, non-arginine methylatable) residues. The significance of the methylation in LANA function was examined in both the isolated form and in the context of the viral genome through the generation of recombinant KSHV. In addition, authentic LANA binding sites on the KSHV episome in naturally infected cells were identified using a whole genome KSHV tiling array. Although mutation of the methylation site resulted in no significant difference in KSHV LANA subcellular localization, we found that the methylation mimetic mutation resulted in augmented histone binding in vitro and increased LANA occupancy at identified LANA target promoters in vivo. Moreover, a cell line carrying the methylation mimetic mutant KSHV showed reduced viral gene expression relative to controls both in latency and in the course of reactivation. These results suggest that residue 20 is important for modulation of a subset of LANA functions and properties of this residue, including the hydrophobic character induced by arginine methylation, may contribute to the observed effects.

AB - The Kaposi sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a multifunctional protein with roles in gene regulation and maintenance of viral latency. Post-translational modification of LANA is important for functional diversification. Here, we report that LANA is subject to arginine methylation by protein arginine methyltransferase 1 in vitro and in vivo. The major arginine methylation site in LANA was mapped to arginine 20. This site was mutated to either phenylalanine (bulky hydrophobic, constitutive methylated mimetic) or lysine (positively charged, non-arginine methylatable) residues. The significance of the methylation in LANA function was examined in both the isolated form and in the context of the viral genome through the generation of recombinant KSHV. In addition, authentic LANA binding sites on the KSHV episome in naturally infected cells were identified using a whole genome KSHV tiling array. Although mutation of the methylation site resulted in no significant difference in KSHV LANA subcellular localization, we found that the methylation mimetic mutation resulted in augmented histone binding in vitro and increased LANA occupancy at identified LANA target promoters in vivo. Moreover, a cell line carrying the methylation mimetic mutant KSHV showed reduced viral gene expression relative to controls both in latency and in the course of reactivation. These results suggest that residue 20 is important for modulation of a subset of LANA functions and properties of this residue, including the hydrophobic character induced by arginine methylation, may contribute to the observed effects.

UR - http://www.scopus.com/inward/record.url?scp=84863115190&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84863115190&partnerID=8YFLogxK

U2 - 10.1074/jbc.M111.289496

DO - 10.1074/jbc.M111.289496

M3 - Article

VL - 287

SP - 5806

EP - 5818

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 8

ER -