DOI: 10.1128/jb.184.2.452-458.2002. Inactivation of the Spirochete <i>recA</i> Gene Results in a Mutant with Low Viability and Irregular Nucleoid Morphology

Inactivation of the Spirochete recA Gene Results in a Mutant with Low Viability and Irregular Nucleoid Morphology

10.1128/jb.184.2.452-458.2002

Crossref journal-article

American Society for Microbiology

Journal of Bacteriology (235)

Abstract

ABSTRACT Recently, we have shown the first evidence for allelic exchange in Leptospira spp. By using the same methodology, the cloned recA of Leptospira biflexa was interrupted by a kanamycin resistance cassette, and the mutated allele was then introduced into the L. biflexa chromosome by homologous recombination. The recA double-crossover mutant showed poor growth in liquid media and was considerably more sensitive to DNA-damaging agents such as mitomycin C and UV light than the wild-type strain. The efficiency of plating of the recA mutant was about 10% of that of the parent strain. In addition, microscopic observation of the L. biflexa recA mutant showed cells that are more elongated than those of the wild-type strain. Fluorescent microscopy of stained cells of the L. biflexa wild-type strain revealed that chromosomal DNA is distributed throughout most of the length of the cell. In contrast, the recA mutant showed aberrant nucleoid morphologies, i.e., DNA is condensed at the midcell. Our data indicate that L. biflexa RecA plays a major role in ensuring cell viability via mechanisms such as DNA repair and, indirectly, active chromosome partitioning.

Bibliography

Kameni, A.-P. T., Couture-Tosi, E., Saint-Girons, I., & Picardeau, M. (2002). Inactivation of the Spirochete recA Gene Results in a Mutant with Low Viability and Irregular Nucleoid Morphology. Journal of Bacteriology, 184(2), 452â458.

Authors 4

Ange-Patricia Tchamedeu Kameni (first)
Evelyne Couture-Tosi (additional)
Isabelle Saint-Girons (additional)
Mathieu Picardeau (additional)

References 25 Referenced 28

10.1128/JB.183.7.2165-2171.2001
Brenot, A., C. Werts, C. Ottone, N. Sertour, N. W. Charon, D. Postic, G. Baranton, and I. Saint Girons. 2001. First evidence for a restriction-modification system in Leptospira sp. FEMS Microbiol. Lett.201:139–143. / FEMS Microbiol. Lett. (2001)
10.1128/jb.118.1.242-249.1974
Casjens, S. 2000. Borrelia genomes in the year 2000. J. Mol. Microbiol. Biotechnol.2:401–410. / J. Mol. Microbiol. Biotechnol. (2000)
Duwat, P., S. D. Ehrlich, and A. Gruss. 1995. The recA gene of Lactococcus lactis: characterization and involvement in oxidative and thermal stress. Mol. Microbiol.17:1121–1131. (10.1111/j.1365-2958.1995.mmi_17061121.x) / Mol. Microbiol. (1995)
Ellinghausen, H. C., and W. G. Mc Cullough. 1965. Nutrition of Leptospira pomona and growth of 13 other serotypes: fractionation of oleic albumin complex and medium bovine albumin and polysorbate 80. Am. J. Vet. Res.26:45–51. / Am. J. Vet. Res. (1965)
10.1038/37551
Fraser, C. M., S. J. Norris, C. M. Weinstock, O. White, G. G. Sutton, R. Dodson, M. Gwinn, E. K. Hickey, R. Clayton, K. A. Ketchum, E. Sodergren, J. M. Hardham, M. P. Mc Leod, S. Salzberg, J. Peterson, H. Khalak, D. Richardson, J. K. Howell, M. Chidambaram, T. Utterback, L. Mc Donald, P. Artiach, C. Bowman, M. D. Cotton, C. Fujii, et al. 1998. Complete genome sequence of Treponema pallidum, the syphilis spirochete. Science281:375–388. (10.1126/science.281.5375.375) / Science (1998)
10.1128/jb.179.7.2228-2237.1997
10.1128/jb.94.1.27-31.1967
10.1128/jb.178.7.1881-1894.1996
Kitten, T., and A. G. Barbour. 1992. The relapsing fever agent Borrelia hermsii has multiple copies of its chromosome and linear plasmids. Genetics132:311–324. (10.1093/genetics/132.2.311) / Genetics (1992)
Kumar, K. K., R. Srivastava, V. B. Sinha, J. Michalski, J. B. Kaper, and B. S. Srivastava. 1994. recA mutations reduce adherence and colonization by classical and El Tor strains of Vibrio cholerae. Microbiology140:1217–1222. (10.1099/13500872-140-5-1217) / Microbiology (1994)
Mackey, B. M., and D. A. Seymour. 1987. The effect of catalase on recovery of heat-injured DNA-repair mutants of Escherichia coli. J. Gen. Microbiol.133:1601–1610. / J. Gen. Microbiol. (1987)
Martin, B., P. Garcia, M. P. Castanié, and J. P. Claverys. 1995. The recA gene of Streptococcus pneumoniae is part of a competence-induced operon and controls lysogenic induction. Mol. Microbiol.15:367–379. (10.1111/j.1365-2958.1995.tb02250.x) / Mol. Microbiol. (1995)
Muth, G., D. Frese, A. Kleber, and W. Wohleben. 1997. Mutational analysis of the Streptomyces lividans recA gene suggests that only mutants with residual activity remain viable. Mol. Gen. Genet.255:420–428. (10.1007/s004380050514) / Mol. Gen. Genet. (1997)
Neidhardt F. C. R. Curtiss III J. L. Ingraham E. C. C. Lin K. B. Low B. Magasanik W. S. Reznikoff M. Riley M. Schaechter and H. E. Umbarger (ed.). 1996. Escherichia coli and Salmonella : cellular and molecular biology vol. 2. ASM Press Washington D.C.
Picardeau, M., A. Brenot, and I. Saint Girons. 2001. First evidence for gene replacement in Leptospiraspp.: tivation of L. biflexa flaB results in nonmotile mutants deficient in endoflagella. Mol. Microbiol.40:189–199. (10.1046/j.1365-2958.2001.02374.x) / Mol. Microbiol. (2001)
Prod’hom, G., B. Lagier, P. Pelicic, A. J. Hance, B. Gicquel, and C. Guilhot. 1998. A reliable amplification technique for the characterization of genomic DNA sequences flanking insertion sequences. FEMS Microbiol. Lett.158:75–81. (10.1111/j.1574-6968.1998.tb12803.x) / FEMS Microbiol. Lett. (1998)
10.1128/JB.182.20.5700-5705.2000
10.1128/JB.183.9.2963-2968.2001
10.1128/JB.182.22.6532-6535.2000
10.1128/aem.57.1.183-189.1991
Trieu-Cuot, P., and P. Courvalin. 1983. Nucleotide sequence of the Streptococcus faecalis plasmid gene encoding the 3′,5′-aminoglycoside phosphotransferase type III. Gene23:331–341. (10.1016/0378-1119(83)90022-7) / Gene (1983)
10.1128/mr.51.2.221-271.1987

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 6:01 a.m.)
Deposited	4 years, 1 month ago (July 29, 2021, 2:05 p.m.)
Indexed	4 months, 3 weeks ago (April 11, 2025, 6:04 a.m.)
Issued	23 years, 7 months ago (Jan. 15, 2002)
Published	23 years, 7 months ago (Jan. 15, 2002)
Published Print	23 years, 7 months ago (Jan. 15, 2002)

Funders 0

None

BibTeX

@article{Kameni_2002, title={Inactivation of the Spirochete recA Gene Results in a Mutant with Low Viability and Irregular Nucleoid Morphology}, volume={184}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.184.2.452-458.2002}, DOI={10.1128/jb.184.2.452-458.2002}, number={2}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Kameni, Ange-Patricia Tchamedeu and Couture-Tosi, Evelyne and Saint-Girons, Isabelle and Picardeau, Mathieu}, year={2002}, month=jan, pages={452–458} }

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