Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Jul;80(14):4422–4426. doi: 10.1073/pnas.80.14.4422

F sex factor of Escherichia coli K-12 codes for a single-stranded DNA binding protein.

A L Kolodkin, M A Capage, E I Golub, K B Low
PMCID: PMC384050  PMID: 6308624

Abstract

In Escherichia coli K-12 strains that carry the mutation ssb-1 in the gene for single-stranded DNA binding protein, the presence of the F sex factor partially reverses the temperature-sensitive growth phenotype caused by the mutation. The region of F (EcoRI fragment 3) responsible for this compensation has been identified and subcloned onto pBR322. A BamHI cleavage site has been found to intersect the essential coding region for this F function. By using this site, mutational blocks in the function have been constructed and used to identify a protein product (Mr approximately 22,000, slightly larger than the E. coli K-12 single-stranded DNA binding protein) which is correlated with the ssb-1-complementing activity. Labeled extracts from maxicells were used to show that this protein binds tightly to single-stranded DNA. The gene on F that codes for this protein is denoted ssf and is located at approximately 55.2 kilobases on the standard map of F, in the region transferred very early during bacterial conjugation.

Full text

PDF
4422

Images in this article

4425Image
on p.4425
4425Image
on p.4425

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Johnson B. F. Suppression of the lexC (ssbA) mutation of Escherichia coli by a mutant of bacteriophage P1. Mol Gen Genet. 1982;186(1):122–126. doi: 10.1007/BF00422923. [DOI] [PubMed] [Google Scholar]
  3. Lane D., Gardner R. C. Second EcoRI fragment of F capable of self-replication. J Bacteriol. 1979 Jul;139(1):141–151. doi: 10.1128/jb.139.1.141-151.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Lieberman H. B., Witkin E. M. Variable expression of the ssb--1 allele in different strains of Escherichia coli K12 and B: differential suppression of its effects on DNA replication, DNA repair and ultraviolet mutagenesis. Mol Gen Genet. 1981;183(2):348–355. doi: 10.1007/BF00270639. [DOI] [PubMed] [Google Scholar]
  5. Lovett M. A., Helinski D. R. Method for the isolation of the replication region of a bacterial replicon: construction of a mini-F'kn plasmid. J Bacteriol. 1976 Aug;127(2):982–987. doi: 10.1128/jb.127.2.982-987.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Meyer R. R., Glassberg J., Kornberg A. An Escherichia coli mutant defective in single-strand binding protein is defective in DNA replication. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1702–1705. doi: 10.1073/pnas.76.4.1702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Molineux I. J., Friedman S., Gefter M. L. Purification and properties of the Escherichia coli deoxyribonucleic acid-unwinding protein. Effects on deoxyribonucleic acid synthesis in vitro. J Biol Chem. 1974 Oct 10;249(19):6090–6098. [PubMed] [Google Scholar]
  9. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sigal N., Delius H., Kornberg T., Gefter M. L., Alberts B. A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3537–3541. doi: 10.1073/pnas.69.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Skurray R. A., Nagaishi H., Clark A. J. Molecular cloning of DNA from F sex factor of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1976 Jan;73(1):64–68. doi: 10.1073/pnas.73.1.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  13. Tessman E. S., Peterson P. K. Suppression of the ssb-1 and ssb-113 mutations of Escherichia coli by a wild-type rep gene, NaCl, and glucose. J Bacteriol. 1982 Nov;152(2):572–583. doi: 10.1128/jb.152.2.572-583.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Timmis K., Cabello F., Cohen S. N. Cloning, isolation, and characterization of replication regions of complex plasmid genomes. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2242–2246. doi: 10.1073/pnas.72.6.2242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Vales L. D., Chase J. W., Murphy J. B. Effect of ssbA1 and lexC113 mutations on lambda prophage induction, bacteriophage growth, and cell survival. J Bacteriol. 1980 Aug;143(2):887–896. doi: 10.1128/jb.143.2.887-896.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Whittier R. F., Chase J. W. DNA repair properties of Escherichia coli tif-1, recAo281 and lexA1 strains deficient in single-strand DNA binding protein. Mol Gen Genet. 1983;190(1):101–111. doi: 10.1007/BF00330330. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES