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. 1993 Sep;175(17):5611–5616. doi: 10.1128/jb.175.17.5611-5616.1993

Mutation in the plasmid pUB110 Rep protein affects termination of rolling circle replication.

V E Bidnenko 1, A Gruss 1, S D Ehrlich 1
PMCID: PMC206618  PMID: 8366045

Abstract

We isolated a mutant of plasmid pUB110 that has the following properties in Bacillus subtilis: (i) it is toxic for recA and add cells, particularly at elevated temperature; (ii) it has a copy number threefold higher than that of the parental plasmid, and the extra copies are present as multimers; and (iii) it can efficiently complement replication of a cmp- satellite plasmid, despite being cmp+. All these properties are due to a single change in the plasmid replication protein, i.e., Gly at position 148 to Glu. These properties of the mutant Rep protein reflect a diminished ability to terminate rolling circle replication. We propose that the Rep protein may have a diminished affinity for the plasmid origin; alternatively, it may be impaired for recognition of the plasmid conformations which distinguish initiation and termination.

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Selected References

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  1. Andreoli P. M., Brandsma R. A., Veltkamp E., Nijkamp H. J. Isolation and characterization of a Clo DF13::Tn901 plasmid mutant with thermosensitive control of DNA replication. J Bacteriol. 1978 Aug;135(2):612–621. doi: 10.1128/jb.135.2.612-621.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  3. Dabert P., Ehrlich S. D., Gruss A. High-molecular-weight linear multimer formation by single-stranded DNA plasmids in Escherichia coli. J Bacteriol. 1992 Jan;174(1):173–178. doi: 10.1128/jb.174.1.173-178.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Doly J., Le Roscouët D., Anagnostopoulos C. Substrate specificity and adenosine triphosphatase activity of the ATP-dependent deoxyribonuclease of Bacillus subtilis. Eur J Biochem. 1981 Mar;114(3):493–499. doi: 10.1111/j.1432-1033.1981.tb05172.x. [DOI] [PubMed] [Google Scholar]
  5. Gennaro M. L., Novick R. P. An enhancer of DNA replication. J Bacteriol. 1988 Dec;170(12):5709–5717. doi: 10.1128/jb.170.12.5709-5717.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gennaro M. L., Novick R. P. cmp, a cis-acting plasmid locus that increases interaction between replication origin and initiator protein. J Bacteriol. 1986 Oct;168(1):160–166. doi: 10.1128/jb.168.1.160-166.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gros M. F., te Riele H., Ehrlich S. D. Replication origin of a single-stranded DNA plasmid pC194. EMBO J. 1989 Sep;8(9):2711–2716. doi: 10.1002/j.1460-2075.1989.tb08412.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gros M. F., te Riele H., Ehrlich S. D. Rolling circle replication of single-stranded DNA plasmid pC194. EMBO J. 1987 Dec 1;6(12):3863–3869. doi: 10.1002/j.1460-2075.1987.tb02724.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gruss A., Ehrlich S. D. Insertion of foreign DNA into plasmids from gram-positive bacteria induces formation of high-molecular-weight plasmid multimers. J Bacteriol. 1988 Mar;170(3):1183–1190. doi: 10.1128/jb.170.3.1183-1190.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gruss A., Ehrlich S. D. The family of highly interrelated single-stranded deoxyribonucleic acid plasmids. Microbiol Rev. 1989 Jun;53(2):231–241. doi: 10.1128/mr.53.2.231-241.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hashimoto T., Sekiguchi M. Isolation of temperature-sensitive mutants of R plasmid by in vitro mutagenesis with hydroxylamine. J Bacteriol. 1976 Sep;127(3):1561–1563. doi: 10.1128/jb.127.3.1561-1563.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Horiuchi K., Ravetch J. V., Zinder N. D. DNA replication of bacteriophage f1 in vivo. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):389–399. doi: 10.1101/sqb.1979.043.01.045. [DOI] [PubMed] [Google Scholar]
  13. Iordanescu S., Projan S. J. Replication termination for staphylococcal plasmids: plasmids pT181 and pC221 cross-react in the termination process. J Bacteriol. 1988 Aug;170(8):3427–3434. doi: 10.1128/jb.170.8.3427-3434.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Koepsel R. R., Murray R. W., Rosenblum W. D., Khan S. A. The replication initiator protein of plasmid pT181 has sequence-specific endonuclease and topoisomerase-like activities. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6845–6849. doi: 10.1073/pnas.82.20.6845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kusano K., Nakayama K., Nakayama H. Plasmid-mediated lethality and plasmid multimer formation in an Escherichia coli recBC sbcBC mutant. Involvement of RecF recombination pathway genes. J Mol Biol. 1989 Oct 20;209(4):623–634. doi: 10.1016/0022-2836(89)90000-4. [DOI] [PubMed] [Google Scholar]
  16. Maciag I. E., Viret J. F., Alonso J. C. Replication and incompatibility properties of plasmid pUB110 in Bacillus subtilis. Mol Gen Genet. 1988 May;212(2):232–240. doi: 10.1007/BF00334690. [DOI] [PubMed] [Google Scholar]
  17. McKenzie T., Hoshino T., Tanaka T., Sueoka N. The nucleotide sequence of pUB110: some salient features in relation to replication and its regulation. Plasmid. 1986 Mar;15(2):93–103. doi: 10.1016/0147-619x(86)90046-6. [DOI] [PubMed] [Google Scholar]
  18. Michel B., Ehrlich S. D. Illegitimate recombination occurs between the replication origin of the plasmid pC194 and a progressing replication fork. EMBO J. 1986 Dec 20;5(13):3691–3696. doi: 10.1002/j.1460-2075.1986.tb04701.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Muller R. E., Ano T., Imanaka T., Aiba S. Complete nucleotide sequences of Bacillus plasmids pUB110dB, pRBH1 and its copy mutants. Mol Gen Genet. 1986 Jan;202(1):169–171. doi: 10.1007/BF00330534. [DOI] [PubMed] [Google Scholar]
  20. Novick R. P. Plasmid incompatibility. Microbiol Rev. 1987 Dec;51(4):381–395. doi: 10.1128/mr.51.4.381-395.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Novick R. P. Staphylococcal plasmids and their replication. Annu Rev Microbiol. 1989;43:537–565. doi: 10.1146/annurev.mi.43.100189.002541. [DOI] [PubMed] [Google Scholar]
  22. Projan S. J., Monod M., Narayanan C. S., Dubnau D. Replication properties of pIM13, a naturally occurring plasmid found in Bacillus subtilis, and of its close relative pE5, a plasmid native to Staphylococcus aureus. J Bacteriol. 1987 Nov;169(11):5131–5139. doi: 10.1128/jb.169.11.5131-5139.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Projan S. J., Novick R. P. cis-inhibitory elements in the pT181 replication system. Plasmid. 1992 Mar;27(2):81–92. doi: 10.1016/0147-619x(92)90009-y. [DOI] [PubMed] [Google Scholar]
  24. Summers D. K., Sherratt D. J. Multimerization of high copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization and stability. Cell. 1984 Apr;36(4):1097–1103. doi: 10.1016/0092-8674(84)90060-6. [DOI] [PubMed] [Google Scholar]
  25. Thomas C. D., Balson D. F., Shaw W. V. In vitro studies of the initiation of staphylococcal plasmid replication. Specificity of RepD for its origin (oriD) and characterization of the Rep-ori tyrosyl ester intermediate. J Biol Chem. 1990 Apr 5;265(10):5519–5530. [PubMed] [Google Scholar]
  26. Viret J. F., Alonso J. C. Generation of linear multigenome-length plasmid molecules in Bacillus subtilis. Nucleic Acids Res. 1987 Aug 25;15(16):6349–6367. doi: 10.1093/nar/15.16.6349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wang P. Z., Henriquez V. B., Projan S. J., Iordanescu S., Novick R. P. The effect of plasmid copy number mutations on pT181 replication initiator protein expression. Plasmid. 1991 May;25(3):198–207. doi: 10.1016/0147-619x(91)90013-m. [DOI] [PubMed] [Google Scholar]
  28. del Solar G. H., de al Campa A. G., Pérez-Martín J., Choli T., Espinosa M. Purification and characterization of RepA, a protein involved in the copy number control of plasmid pLS1. Nucleic Acids Res. 1989 Apr 11;17(7):2405–2420. doi: 10.1093/nar/17.7.2405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. te Riele H., Michel B., Ehrlich S. D. Single-stranded plasmid DNA in Bacillus subtilis and Staphylococcus aureus. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2541–2545. doi: 10.1073/pnas.83.8.2541. [DOI] [PMC free article] [PubMed] [Google Scholar]

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