A recA null mutation may be generated in Streptomyces coelicolor

Tzu-Wen Huang, Hsiuh-Wei Chen

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The recombinase RecA plays a crucial role in homologous recombination and the SOS response in bacteria. Although recA mutants usually are defective in homologous recombination and grow poorly, they nevertheless can be isolated in almost all bacteria. Previously, considerable difficulties were experienced by several laboratories in generating recA null mutations in Streptomyces, and the only recA null mutants isolated (from Streptomyces lividans) appeared to be accompanied by a suppressing mutation. Using gene replacement mediated by Escherichia coli-Streptomyces conjugation, we generated recA null mutations in a series of Streptomyces coelicolor A3 (2) strains. These recA mutants were very sensitive to mitomycin C but only moderately sensitive to UV irradiation, and the UV survival curves showed wide shoulders, reflecting the presence of a recA-independent repair pathway. The mutants segregated minute colonies with low viability during growth and produced more anucleate spores than the wild type. Some crosses between pairs of recA null mutants generated no detectable recombinants, showing for the first time that conjugal recombination in S. coelicolor is recA mediated, but other mutants retained the ability to undergo recombination. The nature of this novel recombination activity is unknown. Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Original languageEnglish
Pages (from-to)6771-6779
Number of pages9
JournalJournal of Bacteriology
Volume188
Issue number19
DOIs
Publication statusPublished - 2006
Externally publishedYes

Fingerprint

Streptomyces coelicolor
Genetic Recombination
Homologous Recombination
Streptomyces
Mutation
Streptomyces lividans
Bacteria
Recombinases
Mitomycin
Microbiology
Spores
Escherichia coli
Growth
Genes

Keywords

  • gene product
  • mitomycin C
  • mutant protein
  • RecA protein
  • recombinase
  • article
  • bacterial gene
  • bacterial genetics
  • bacterial growth
  • bacterial mutation
  • bacterial spore
  • bacterial strain
  • bacterial survival
  • bacterium colony
  • bacterium conjugation
  • bacterium isolation
  • bacterium mutant
  • cell viability
  • controlled study
  • DNA repair
  • drug sensitivity
  • Escherichia coli
  • gene mutation
  • homologous recombination
  • nonhuman
  • null allele
  • priority journal
  • radiosensitivity
  • RecA gene
  • Streptomyces coelicolor
  • Streptomyces lividans
  • ultraviolet radiation
  • Anti-Bacterial Agents
  • Conjugation, Genetic
  • Gene Deletion
  • Mitomycin
  • Rec A Recombinases
  • Recombination, Genetic
  • Ultraviolet Rays
  • Streptomyces

Cite this

A recA null mutation may be generated in Streptomyces coelicolor. / Huang, Tzu-Wen; Chen, Hsiuh-Wei.

In: Journal of Bacteriology, Vol. 188, No. 19, 2006, p. 6771-6779.

Research output: Contribution to journalArticle

Huang, Tzu-Wen ; Chen, Hsiuh-Wei. / A recA null mutation may be generated in Streptomyces coelicolor. In: Journal of Bacteriology. 2006 ; Vol. 188, No. 19. pp. 6771-6779.
@article{18e11174de1c41de960713c3964b12a0,
title = "A recA null mutation may be generated in Streptomyces coelicolor",
abstract = "The recombinase RecA plays a crucial role in homologous recombination and the SOS response in bacteria. Although recA mutants usually are defective in homologous recombination and grow poorly, they nevertheless can be isolated in almost all bacteria. Previously, considerable difficulties were experienced by several laboratories in generating recA null mutations in Streptomyces, and the only recA null mutants isolated (from Streptomyces lividans) appeared to be accompanied by a suppressing mutation. Using gene replacement mediated by Escherichia coli-Streptomyces conjugation, we generated recA null mutations in a series of Streptomyces coelicolor A3 (2) strains. These recA mutants were very sensitive to mitomycin C but only moderately sensitive to UV irradiation, and the UV survival curves showed wide shoulders, reflecting the presence of a recA-independent repair pathway. The mutants segregated minute colonies with low viability during growth and produced more anucleate spores than the wild type. Some crosses between pairs of recA null mutants generated no detectable recombinants, showing for the first time that conjugal recombination in S. coelicolor is recA mediated, but other mutants retained the ability to undergo recombination. The nature of this novel recombination activity is unknown. Copyright {\circledC} 2006, American Society for Microbiology. All Rights Reserved.",
keywords = "gene product, mitomycin C, mutant protein, RecA protein, recombinase, article, bacterial gene, bacterial genetics, bacterial growth, bacterial mutation, bacterial spore, bacterial strain, bacterial survival, bacterium colony, bacterium conjugation, bacterium isolation, bacterium mutant, cell viability, controlled study, DNA repair, drug sensitivity, Escherichia coli, gene mutation, homologous recombination, nonhuman, null allele, priority journal, radiosensitivity, RecA gene, Streptomyces coelicolor, Streptomyces lividans, ultraviolet radiation, Anti-Bacterial Agents, Conjugation, Genetic, Gene Deletion, Mitomycin, Rec A Recombinases, Recombination, Genetic, Ultraviolet Rays, Streptomyces",
author = "Tzu-Wen Huang and Hsiuh-Wei Chen",
note = "Export Date: 13 April 2016 CODEN: JOBAA 通訊地址: Chen, C.W.; Faculty of Life Sciences, Institute of Genome Sciences, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan; 電子郵件: cwchen@ym.edu.tw 化學物質/CAS: mitomycin C, 50-07-7, 74349-48-7; RecA protein, 73177-10-3; Anti-Bacterial Agents; Mitomycin, 50-07-7; Rec A Recombinases, EC 2.7.7.- 參考文獻: Ahel, I., Mikoc, A., Gamulin, V., recA gene expression in a streptomycete is mediated by the unusual C-terminus of RecA protein (2005) FEMS Microbiol. Lett., 248, pp. 119-124; Aigle, B., Holl, A.C., Angulo, J.F., Leblond, P., Decaris, B., Characterization of two Streptomyces ambofaciens recA mutants: Identification of the RecA protein by immunoblotting (1997) FEMS Microbiol. Lett., 149, pp. 181-187; Bentley, S.D., Chater, K.F., Cerde{\~n}o-T{\'a}rraga, A.-M., Challis, G.L., Thomson, N.R., James, K.D., Harris, D.E., Hopwood, D.A., Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2) (2002) Nature, 417, pp. 141-147; Capaldo, F.N., Barbour, S.D., DNA content, synthesis and integrity in dividing and non-dividing cells of rec - strains of Escherichia coli K12 (1975) J. Mol. Biol., 91, pp. 53-66; Capaldo, F.N., Ramsey, G., Barbour, S.D., Analysis of the growth of recombination-deficient strains of Escherichia coli K-12 (1974) J. Bacteriol., 118, pp. 242-249; Casjens, S., Prophages and bacterial genomics: What have we learned so far? (2003) Mol. Microbiol., 49, pp. 277-300; Chen, C.W., Complications and implications of linear bacterial chromosomes (1996) Trends Genet., 12, pp. 192-196; Chen, C.W., Huang, C.-H., Lee, H.-H., Tsai, H.-H., Kirby, R., Once the circle has been broken: Dynamics and evolution of Streptomyces chromosomes (2002) Trends Genet., 18, pp. 522-529; Clark, A.J., Margulies, A.D., Isolation and characterization of recombination-deficient mutants of Escherichia coli K12 (1965) Proc. Natl. Acad. Sci. USA, 53, pp. 451-459; Gust, B., Challis, G.L., Fowler, K., Kieser, T., Chater, K.F., PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin (2003) Proc. Natl. Acad. Sci. USA, 100, pp. 1541-1546; Haefner, K., Spontaneous lethal sectoring, a further feature of Escherichia coli strains deficient in the function of rec and uvr genes (1968) J. Bacteriol., 96, pp. 652-659; Harold, R., Hopwood, D., Ultraviolet-sensitive mutants of Streptomyces coelicolor (1970) Mutat. Res., 10, pp. 427-438; Hertman, I.M., Isolation and characterization of a recombination-deficient Hfr strain (1967) J. Bacteriol., 93, pp. 580-583; Hopwood, D.A., Forty years of genetics with Streptomyces: From in vivo through in vitro to in silico (1999) Microbiology, 145, pp. 2183-2202; Hopwood, D.A., Soil to genomics: The Streptomyces chromosome Annu. Rev. Genet., , in press; Hopwood, D.A., Chafer, K.F., Dowding, J.E., Vivian, A., Advances in Streptomyces coelicolor genetics (1973) Bacteriol. Rev., 37, pp. 371-405; Hopwood, D.A., Harold, R.J., Vivian, A., Ferguson, H.M., A new kind of fertility variant in Streptomyces coelicolor (1969) Genetics, 62, pp. 461-477; Hopwood, D.A., Kieser, T., Wright, H.M., Bibb, M.J., Plasmids, recombination, and chromosomal mapping in Streptomyces lividans 66 (1983) J. Gen. Microbiol., 129, pp. 2257-2269; Ikeda, H., Ishikawa, J., Hanamoto, A., Shinose, M., Kikuchi, H., Shiba, T., Sakaki, Y., Omura, S., Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis (2003) Nat. Biotechnol., 21, pp. 526-531; Kieser, T., Bibb, M., Buttner, M.J., Chater, K.F., Hopwood, D.A., (2000) Practical Streptomyces Genetics, , The John Innes Foundation, Norwich, United Kingdom; Kim, H.J., Calcutt, M.J., Schmidt, F.J., Chater, K.F., Partitioning of the linear chromosome during sporulation of Streptomyces coelicolor A3(2) involves an oriC-linked parAB locus (2000) J. Bacteriol., 182, pp. 1313-1320; Kuzminov, A., Recombinational repair of DNA damage in Escherichia coli and bacteriophage λ (1999) Microbiol. Mol. Biol. Rev., 63, pp. 751-813; Lee, C.-C., (1997) Construction and Characterization of RecF Mutants of Streptomyces Lividans ZX7, , M.S. thesis. National Yang-Ming University, Taipei, Taiwan; Lin, Y.-S., Kieser, H.M., Hopwood, D.A., Chen, C.W., The chromosomal DNA of Streptomyces lividans 66 is linear (1993) Mol. Microbiol., 10, pp. 923-933; Mikoč, A., Ahel, I., Gamulin, V., Construction and characterization of a Streptomyces rimosus recA mutant: The RecA-deficient strain remains viable (2000) Mol. Gen. Genet., 264, pp. 227-232; Mikoč, A., Vujaklija, D., Gamulin, V., The recA gene from Streptomyces rimosus R6: Sequence and expression in Escherichia coli (1997) Res. Microbiol., 148, pp. 397-403; Muth, G., Frese, D., Kleber, A., Wohlleben, W., Mutational analysis of the Streptomyces lividans recA gene suggests that only mutants with residual activity remain viable (1997) Mol. Gen. Genet., 255, pp. 420-428; Norioka, N., Hsu, M.Y., Inouye, S., Inouye, M., Two recA genes in Myxococcus xanthus (1995) J. Bacteriol., 177, pp. 4179-4182; Paget, M.S., Chamberlin, L., Atrih, A., Foster, S.J., Buttner, M.J., Evidence that the extracytoplasmic function sigma factor σ E is required for normal cell wall structure in Streptomyces coelicolor A3(2) (1999) J. Bacteriol., 181, pp. 204-211; Possoz, C., Ribard, C., Gagnat, J., Pernodet, J.L., Guerineau, M., The integrative element pSAM2 from Streptomyces: Kinetics and mode of conjugal transfer (2001) Mol. Microbiol., 42, pp. 159-166; Putteet-Driver, A.D., Zhong, J., Harbour, A.G., Transgenic expression of RecA of the spirochetes Borrelia burgdorferi and Borrelia hermsii in Escherichia coli revealed differences in DNA repair and recombination phenotypes (2004) J. Bacteriol., 186, pp. 2266-2274; Qin, Z., Cohen, S.N., Replication at the telomeres of the Streptomyces linear plasmid pSLA2 (1998) Mol. Microbiol., 28, pp. 893-904; Redenbach, M., Kieser, H.M., Denapaite, D., Eichner, A., Cullum, J., Kinashi, H., Hopwood, D.A., A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome (1996) Mol. Microbiol., 21, pp. 77-96; Rocha, E.P., Cornet, E., Michel, B., Comparative and evolutionary analysis of the bacterial homologous recombination systems (2005) PLoS Genet., 1, pp. e15; Sambrook, J., MacCallum, P., Russel, D., (2001) Molecular Cloning, 3rd Ed., , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y; Vierling, S., Weber, T., Wohlleben, W., Muth, G., Evidence that an additional mutation is required to tolerate insertional inactivation of the Streptomyces lividans recA gene (2001) J. Bacteriol., 183, pp. 4374-4381; Wang, S.-J., Chang, H.-M., Lin, Y.-S., Huang, C.-H., Chen, C.W., Streptomyces genomes: Circular genetic maps from the linear chromosomes (1999) Microbiology, 145, pp. 2209-2220; Weaver, D., Karoonuthaisiri, N., Tsai, H.H., Huang, C.H., Ho, M.L., Gai, S., Patel, K.G., Kao, C.M., Genome plasticity in Streptomyces: Identification of 1 Mb TIR in the S. coelicolor A3(2) chromosome (2004) Mol. Microbiol., 51, pp. 1530-1550; Weller, G.R., Kysela, B., Roy, R., Tonkin, L.M., Scanlan, E., Della, M., Devine, S.K., Doherty, A.J., Identification of a DNA nonhomologous end-joining complex in bacteria (2002) Science, 297, pp. 1686-1689",
year = "2006",
doi = "10.1128/JB.00951-06",
language = "English",
volume = "188",
pages = "6771--6779",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "19",

}

TY - JOUR

T1 - A recA null mutation may be generated in Streptomyces coelicolor

AU - Huang, Tzu-Wen

AU - Chen, Hsiuh-Wei

N1 - Export Date: 13 April 2016 CODEN: JOBAA 通訊地址: Chen, C.W.; Faculty of Life Sciences, Institute of Genome Sciences, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan; 電子郵件: cwchen@ym.edu.tw 化學物質/CAS: mitomycin C, 50-07-7, 74349-48-7; RecA protein, 73177-10-3; Anti-Bacterial Agents; Mitomycin, 50-07-7; Rec A Recombinases, EC 2.7.7.- 參考文獻: Ahel, I., Mikoc, A., Gamulin, V., recA gene expression in a streptomycete is mediated by the unusual C-terminus of RecA protein (2005) FEMS Microbiol. Lett., 248, pp. 119-124; Aigle, B., Holl, A.C., Angulo, J.F., Leblond, P., Decaris, B., Characterization of two Streptomyces ambofaciens recA mutants: Identification of the RecA protein by immunoblotting (1997) FEMS Microbiol. Lett., 149, pp. 181-187; Bentley, S.D., Chater, K.F., Cerdeño-Tárraga, A.-M., Challis, G.L., Thomson, N.R., James, K.D., Harris, D.E., Hopwood, D.A., Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2) (2002) Nature, 417, pp. 141-147; Capaldo, F.N., Barbour, S.D., DNA content, synthesis and integrity in dividing and non-dividing cells of rec - strains of Escherichia coli K12 (1975) J. Mol. Biol., 91, pp. 53-66; Capaldo, F.N., Ramsey, G., Barbour, S.D., Analysis of the growth of recombination-deficient strains of Escherichia coli K-12 (1974) J. Bacteriol., 118, pp. 242-249; Casjens, S., Prophages and bacterial genomics: What have we learned so far? (2003) Mol. Microbiol., 49, pp. 277-300; Chen, C.W., Complications and implications of linear bacterial chromosomes (1996) Trends Genet., 12, pp. 192-196; Chen, C.W., Huang, C.-H., Lee, H.-H., Tsai, H.-H., Kirby, R., Once the circle has been broken: Dynamics and evolution of Streptomyces chromosomes (2002) Trends Genet., 18, pp. 522-529; Clark, A.J., Margulies, A.D., Isolation and characterization of recombination-deficient mutants of Escherichia coli K12 (1965) Proc. Natl. Acad. Sci. USA, 53, pp. 451-459; Gust, B., Challis, G.L., Fowler, K., Kieser, T., Chater, K.F., PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin (2003) Proc. Natl. Acad. Sci. USA, 100, pp. 1541-1546; Haefner, K., Spontaneous lethal sectoring, a further feature of Escherichia coli strains deficient in the function of rec and uvr genes (1968) J. Bacteriol., 96, pp. 652-659; Harold, R., Hopwood, D., Ultraviolet-sensitive mutants of Streptomyces coelicolor (1970) Mutat. Res., 10, pp. 427-438; Hertman, I.M., Isolation and characterization of a recombination-deficient Hfr strain (1967) J. Bacteriol., 93, pp. 580-583; Hopwood, D.A., Forty years of genetics with Streptomyces: From in vivo through in vitro to in silico (1999) Microbiology, 145, pp. 2183-2202; Hopwood, D.A., Soil to genomics: The Streptomyces chromosome Annu. Rev. Genet., , in press; Hopwood, D.A., Chafer, K.F., Dowding, J.E., Vivian, A., Advances in Streptomyces coelicolor genetics (1973) Bacteriol. Rev., 37, pp. 371-405; Hopwood, D.A., Harold, R.J., Vivian, A., Ferguson, H.M., A new kind of fertility variant in Streptomyces coelicolor (1969) Genetics, 62, pp. 461-477; Hopwood, D.A., Kieser, T., Wright, H.M., Bibb, M.J., Plasmids, recombination, and chromosomal mapping in Streptomyces lividans 66 (1983) J. Gen. Microbiol., 129, pp. 2257-2269; Ikeda, H., Ishikawa, J., Hanamoto, A., Shinose, M., Kikuchi, H., Shiba, T., Sakaki, Y., Omura, S., Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis (2003) Nat. Biotechnol., 21, pp. 526-531; Kieser, T., Bibb, M., Buttner, M.J., Chater, K.F., Hopwood, D.A., (2000) Practical Streptomyces Genetics, , The John Innes Foundation, Norwich, United Kingdom; Kim, H.J., Calcutt, M.J., Schmidt, F.J., Chater, K.F., Partitioning of the linear chromosome during sporulation of Streptomyces coelicolor A3(2) involves an oriC-linked parAB locus (2000) J. Bacteriol., 182, pp. 1313-1320; Kuzminov, A., Recombinational repair of DNA damage in Escherichia coli and bacteriophage λ (1999) Microbiol. Mol. Biol. Rev., 63, pp. 751-813; Lee, C.-C., (1997) Construction and Characterization of RecF Mutants of Streptomyces Lividans ZX7, , M.S. thesis. National Yang-Ming University, Taipei, Taiwan; Lin, Y.-S., Kieser, H.M., Hopwood, D.A., Chen, C.W., The chromosomal DNA of Streptomyces lividans 66 is linear (1993) Mol. Microbiol., 10, pp. 923-933; Mikoč, A., Ahel, I., Gamulin, V., Construction and characterization of a Streptomyces rimosus recA mutant: The RecA-deficient strain remains viable (2000) Mol. Gen. Genet., 264, pp. 227-232; Mikoč, A., Vujaklija, D., Gamulin, V., The recA gene from Streptomyces rimosus R6: Sequence and expression in Escherichia coli (1997) Res. Microbiol., 148, pp. 397-403; Muth, G., Frese, D., Kleber, A., Wohlleben, W., Mutational analysis of the Streptomyces lividans recA gene suggests that only mutants with residual activity remain viable (1997) Mol. Gen. Genet., 255, pp. 420-428; Norioka, N., Hsu, M.Y., Inouye, S., Inouye, M., Two recA genes in Myxococcus xanthus (1995) J. Bacteriol., 177, pp. 4179-4182; Paget, M.S., Chamberlin, L., Atrih, A., Foster, S.J., Buttner, M.J., Evidence that the extracytoplasmic function sigma factor σ E is required for normal cell wall structure in Streptomyces coelicolor A3(2) (1999) J. Bacteriol., 181, pp. 204-211; Possoz, C., Ribard, C., Gagnat, J., Pernodet, J.L., Guerineau, M., The integrative element pSAM2 from Streptomyces: Kinetics and mode of conjugal transfer (2001) Mol. Microbiol., 42, pp. 159-166; Putteet-Driver, A.D., Zhong, J., Harbour, A.G., Transgenic expression of RecA of the spirochetes Borrelia burgdorferi and Borrelia hermsii in Escherichia coli revealed differences in DNA repair and recombination phenotypes (2004) J. Bacteriol., 186, pp. 2266-2274; Qin, Z., Cohen, S.N., Replication at the telomeres of the Streptomyces linear plasmid pSLA2 (1998) Mol. Microbiol., 28, pp. 893-904; Redenbach, M., Kieser, H.M., Denapaite, D., Eichner, A., Cullum, J., Kinashi, H., Hopwood, D.A., A set of ordered cosmids and a detailed genetic and physical map for the 8 Mb Streptomyces coelicolor A3(2) chromosome (1996) Mol. Microbiol., 21, pp. 77-96; Rocha, E.P., Cornet, E., Michel, B., Comparative and evolutionary analysis of the bacterial homologous recombination systems (2005) PLoS Genet., 1, pp. e15; Sambrook, J., MacCallum, P., Russel, D., (2001) Molecular Cloning, 3rd Ed., , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y; Vierling, S., Weber, T., Wohlleben, W., Muth, G., Evidence that an additional mutation is required to tolerate insertional inactivation of the Streptomyces lividans recA gene (2001) J. Bacteriol., 183, pp. 4374-4381; Wang, S.-J., Chang, H.-M., Lin, Y.-S., Huang, C.-H., Chen, C.W., Streptomyces genomes: Circular genetic maps from the linear chromosomes (1999) Microbiology, 145, pp. 2209-2220; Weaver, D., Karoonuthaisiri, N., Tsai, H.H., Huang, C.H., Ho, M.L., Gai, S., Patel, K.G., Kao, C.M., Genome plasticity in Streptomyces: Identification of 1 Mb TIR in the S. coelicolor A3(2) chromosome (2004) Mol. Microbiol., 51, pp. 1530-1550; Weller, G.R., Kysela, B., Roy, R., Tonkin, L.M., Scanlan, E., Della, M., Devine, S.K., Doherty, A.J., Identification of a DNA nonhomologous end-joining complex in bacteria (2002) Science, 297, pp. 1686-1689

PY - 2006

Y1 - 2006

N2 - The recombinase RecA plays a crucial role in homologous recombination and the SOS response in bacteria. Although recA mutants usually are defective in homologous recombination and grow poorly, they nevertheless can be isolated in almost all bacteria. Previously, considerable difficulties were experienced by several laboratories in generating recA null mutations in Streptomyces, and the only recA null mutants isolated (from Streptomyces lividans) appeared to be accompanied by a suppressing mutation. Using gene replacement mediated by Escherichia coli-Streptomyces conjugation, we generated recA null mutations in a series of Streptomyces coelicolor A3 (2) strains. These recA mutants were very sensitive to mitomycin C but only moderately sensitive to UV irradiation, and the UV survival curves showed wide shoulders, reflecting the presence of a recA-independent repair pathway. The mutants segregated minute colonies with low viability during growth and produced more anucleate spores than the wild type. Some crosses between pairs of recA null mutants generated no detectable recombinants, showing for the first time that conjugal recombination in S. coelicolor is recA mediated, but other mutants retained the ability to undergo recombination. The nature of this novel recombination activity is unknown. Copyright © 2006, American Society for Microbiology. All Rights Reserved.

AB - The recombinase RecA plays a crucial role in homologous recombination and the SOS response in bacteria. Although recA mutants usually are defective in homologous recombination and grow poorly, they nevertheless can be isolated in almost all bacteria. Previously, considerable difficulties were experienced by several laboratories in generating recA null mutations in Streptomyces, and the only recA null mutants isolated (from Streptomyces lividans) appeared to be accompanied by a suppressing mutation. Using gene replacement mediated by Escherichia coli-Streptomyces conjugation, we generated recA null mutations in a series of Streptomyces coelicolor A3 (2) strains. These recA mutants were very sensitive to mitomycin C but only moderately sensitive to UV irradiation, and the UV survival curves showed wide shoulders, reflecting the presence of a recA-independent repair pathway. The mutants segregated minute colonies with low viability during growth and produced more anucleate spores than the wild type. Some crosses between pairs of recA null mutants generated no detectable recombinants, showing for the first time that conjugal recombination in S. coelicolor is recA mediated, but other mutants retained the ability to undergo recombination. The nature of this novel recombination activity is unknown. Copyright © 2006, American Society for Microbiology. All Rights Reserved.

KW - gene product

KW - mitomycin C

KW - mutant protein

KW - RecA protein

KW - recombinase

KW - article

KW - bacterial gene

KW - bacterial genetics

KW - bacterial growth

KW - bacterial mutation

KW - bacterial spore

KW - bacterial strain

KW - bacterial survival

KW - bacterium colony

KW - bacterium conjugation

KW - bacterium isolation

KW - bacterium mutant

KW - cell viability

KW - controlled study

KW - DNA repair

KW - drug sensitivity

KW - Escherichia coli

KW - gene mutation

KW - homologous recombination

KW - nonhuman

KW - null allele

KW - priority journal

KW - radiosensitivity

KW - RecA gene

KW - Streptomyces coelicolor

KW - Streptomyces lividans

KW - ultraviolet radiation

KW - Anti-Bacterial Agents

KW - Conjugation, Genetic

KW - Gene Deletion

KW - Mitomycin

KW - Rec A Recombinases

KW - Recombination, Genetic

KW - Ultraviolet Rays

KW - Streptomyces

U2 - 10.1128/JB.00951-06

DO - 10.1128/JB.00951-06

M3 - Article

VL - 188

SP - 6771

EP - 6779

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 19

ER -