1887

Abstract

The gene coding for the homologue of LexA has been cloned and sequenced. Amino acids required for autocatalytic cleavage are conserved, whereas those important for specific DNA binding are not, when compared with LexA. The transcriptional start site was mapped and a DNA sequence motif was identified which resembled the consensus Cheo box sequence involved in the regulation of DNA-damage-inducible genes in . The LexA protein was overexpressed in and purified by means of a His tag. The purifed LexA was shown to bind to the Cheo box sequence found upstream of its own gene.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-3-929
1997-03-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/3/mic-143-3-929.html?itemId=/content/journal/micro/10.1099/00221287-143-3-929&mimeType=html&fmt=ahah

References

  1. Brent R., Ptashne M. 1981; Mechanism of action of the lexA gene product.. Proc Natl Acad Sci USA 78:4204–4208
    [Google Scholar]
  2. Cheo D.L, Bayles K.W., Yasbin R.E. 1991; Cloning and characterisation of DNA damage inducible promoter regions from Bacillus subtilis.. J Bacteriol 173:1696–1703
    [Google Scholar]
  3. Davis E.O., Sedgwick S.G., Colston M.J. 1991; Novel structure of the recA locus of Mycobacterium tuberculosis implies processing of the gene product.. J Bacteriol 173:5653–5662
    [Google Scholar]
  4. Davis E.O., Thangaraj H.S., Brooks P.C., Colston M.J. 1994; Evidence of selection for protein introns in the RecAs of pathogenic mycobacteria.. EMBO J 13:699–703
    [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Res 12: 387–395
    [Google Scholar]
  6. Fogh R.H., Ottleben G., Ruterjans H., Schnarr M., Boelens R., Kaptein R. 1994; Solution structure of the LexA repressor DNA binding domain determined by H+ NMR spectroscopy.. EMBO J 13: 3936–3944
    [Google Scholar]
  7. Garriga X., Calero S., Barbe J. 1992; Nucleotide sequence analysis and comparison of the lexA genes from Salmonella typbimurium, Erwinia carotovora, Pseudomonas aeruginosa and Pseudomonas putida.. Mol Gen Genet 236:125–134
    [Google Scholar]
  8. Gonzalez-y-Merchand J.A., Colston M.J., Cox R.A. 1996; The rRNA operons of Mycobacterium smegmatis and Myco¬bacterium tuberculosis: comparison of promoter elements and of neighbouring upstream genes.. Microbiology 142:667–674
    [Google Scholar]
  9. Holmes D.S., Quigley M. 1981; A rapid boiling method for the preparation of bacterial plasmids.. Anal Biochem 114:193–197
    [Google Scholar]
  10. Horii T., Ogawa T., Nakatani T., Hase T., Matsubara H., Ogawa H. 1981a; Regulation of SOS functions: purification of E. coli LexA protein and determination of its specific site cleaved by the RecA protein.. Cell 27:515–522
    [Google Scholar]
  11. Horii T., Ogawa T., Ogawa H. 1981b; Nucleotide sequence of the lexA gene of E. coli.. Cell 23:689–697
    [Google Scholar]
  12. Knegtel R.M.A., Fogh R.H., Ottleben G., Ruterjans H., Dumoulin P., Schnarr M., Boelens R., Kaptein R. 1995; A model for the LexA repressor DNA complex.. Proteins 21:226–236
    [Google Scholar]
  13. Kochi A. 1991; The global tuberculosis situation and the new control strategy of the World Health Organisation.. Tubercle 72:1–6
    [Google Scholar]
  14. Little J.W. 1984; Autodigestion of lexA and phage lambda repressors.. Proc Natl Acad Sci USA 81:1375–1379
    [Google Scholar]
  15. Little J.W. 1991; Mechanism of specific LexA cleavage: autodigestion and the role of RecA coprotease.. Biochimie 73:411–422
    [Google Scholar]
  16. Little J.W., Mount D.W. 1982; The SOS regulatory system of Escherichia coli.. Cell 29:11–22
    [Google Scholar]
  17. Little J.W., Kim B., Roland K.L, Smith M.H., Lin L.L., Slilaty S.N. 1994; Cleavage of LexA repressor.. Methods Enzymol 244:266–284
    [Google Scholar]
  18. Markham B.E., Little J.W. 1981; Nucleotide sequence of the lexA gene of Escherichia coli K-12.. Nucleic Acids Res 9:4149–4161
    [Google Scholar]
  19. Mead D.A., Szczesna-Skorupa E., Kemper B. 1987; Singlestranded DNA ‘blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering.. Protein Eng 1:67–74
    [Google Scholar]
  20. Miller R.V., Kokjohn T.A. 1990; General microbiology of recA: environmental and evolutionary significance.. Annu Rev Microbiol 44:365–394
    [Google Scholar]
  21. Mustard J.A., Thliveris A.T., Mount D.W. 1992; Sequence of the Salmonella typhimurium LT2 lexA gene and its regulatory region.. Nucleic Acids Res 20:1813
    [Google Scholar]
  22. Philipp W.J., Poulet S., Eiglmeier K., Pascopella L., Balasubramanian V., Heym B., Bergh S., Bloom B.R., Jacobs W.R., Cole S.T. 1996; An integrated map of the genome of the tubercle bacillus, Mycobacterium tuberculosis H37Rv, and comparison with Mycobacterium leprae.. Proc Natl Acad Sci USA 93:3132–3137
    [Google Scholar]
  23. Raymond-Denise A., Guillen N. 1991; Identification of dinR, a DNA damage-inducible regulator gene of Bacillus subtilis. . J Bacteriol 173:7084–7091
    [Google Scholar]
  24. Riera J., Barbe J. 1993; Sequence of the Providencia rettgeri lexA gene and its control region.. Nucleic Acids Res 21:2256
    [Google Scholar]
  25. Riera J., Barbe J. 1995; Cloning, sequence and regulation of expression of the lexA gene of Aeromonas hydrophila.. Gene 154:71–75
    [Google Scholar]
  26. Rosenberg M., Court D. 1979; Regulatory sequences involved in the promotion and termination of RNA transcription.. Annu Rev Genet 13:319–353
    [Google Scholar]
  27. Sambrook J., Fritsch E.F., Maniatias T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Sarkis G.J., Jacobs W.R., Hatfull G.F. 1995; L5 luciferase reporter mycobacteriophages: a sensitive tool for the detection and assay of live mycobacteria.. Mol Microbiol 15:1055–1067
    [Google Scholar]
  29. Slilaty S.N., Little J.W. 1987; Lysine-156 and serine-119 are required for LexA repressor cleavage: a possible mechanism.. Proc Natl Acad Sci USA 84:3987–3991
    [Google Scholar]
  30. Studier F.W., Rosenberg A.H., Dunn J.J., Dubendorff J.W. 1990; Use of T7 RNA polymerase to direct expression of cloned genes.. Methods Enzymol 185:60–89
    [Google Scholar]
  31. Suzuki M., Yagi N., Gerstein M. 1995; DNA recognition and superstructure formation by helix-turn-helix proteins.. Protein Eng 8:329–338
    [Google Scholar]
  32. Thliveris A.T., Mount D.W. 1992; Genetic identification of the DNA binding domain of Escherichia coli LexA protein.. Proc Natl Acad Sci USA 89:4500–4504
    [Google Scholar]
  33. Thliveris A.T., Little J.W., Mount D.W. 1991; Represssion of the E. coli recA gene requires at least two LexA protein monomers.. Biochimie 73:449–455
    [Google Scholar]
  34. Timm J., Perilli M.G., Duez C., Trias J., Orefici G., Fattorini L., Amicosante G., Oratore A., Joris B., Frere J.M., Pugsley A.P., Gicquel B. 1994; Transcription and expression analysis, using lacZ and phoA gene fusions, of Mycobacterium fortuitum βLlactamase genes cloned from a natural isolate and a high-level β-lactamase producer.. Mol Microbiol 12:491–504
    [Google Scholar]
  35. Walker G.C. 1984; Mutagenesis and inducible responses to deoxyribonucleic acid damage in E. coli.. Microbiol Rev 48:60–93
    [Google Scholar]
  36. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors.. Gene 33:103–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-3-929
Loading
/content/journal/micro/10.1099/00221287-143-3-929
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error