Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Nov;172(11):6551–6556. doi: 10.1128/jb.172.11.6551-6556.1990

Physical analysis of phr gene transcription in Escherichia coli K-12.

M C Lorence 1, S D Maika 1, C S Rupert 1
PMCID: PMC526844  PMID: 1699932

Abstract

The phr gene of Escherichia coli K-12 encodes the light-dependent, DNA repair enzyme photolyase, which removes UV light-induced pyrimidine dimers from cellular DNA. From Southern hybridization analysis of several strains containing successively extended phr deletions, we have determined the direction of transcription of the phr gene on the E. coli K-12 chromosome. Northern (RNA) hybridization analysis suggests that the phr gene is cotranscribed with a previously identified gene of unknown function (orf169) into two messages of different lengths. S1 nuclease mapping analysis indicates that the two transcripts share a single termination site but initiate at two different sites. Finally, we have determined that the presence of orf169 is not necessary for phr gene activity in vivo.

Full text

PDF
6554

Images in this article

6553Image
on p.6553
6553Image
on p.6553
6554Image
on p.6554
6554Image
on p.6554

Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Harm W. Biological determination of the germicidal activity of sunlight. Radiat Res. 1969 Oct;40(1):63–69. [PubMed] [Google Scholar]
  8. Harm W., Harm H., Rupert C. S. Analysis of photoenzymatic repair of UV lesions in DNA by single light flashes. II. In vivo studies with Escherichia coli cells and bacteriophage. Mutat Res. 1968 Nov-Dec;6(3):371–385. doi: 10.1016/0027-5107(68)90054-7. [DOI] [PubMed] [Google Scholar]
  9. Horowitz H., Platt T. Initiation in vivo at the internal trp p2 promoter of Escherichia coli. J Biol Chem. 1983 Jul 10;258(13):7890–7893. [PubMed] [Google Scholar]
  10. Ihara M., Yamamoto K., Ohnishi T. Induction of phr gene expression by irradiation of ultraviolet light in Escherichia coli. Mol Gen Genet. 1987 Aug;209(1):200–202. doi: 10.1007/BF00329860. [DOI] [PubMed] [Google Scholar]
  11. JAGGER J. A small and inexpensive ultraviolet dose-rate meter useful in biological experiements. Radiat Res. 1961 Apr;14:394–403. [PubMed] [Google Scholar]
  12. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  13. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  14. Locker J. Analytical and preparative electrophoresis of RNA in agarose-urea. Anal Biochem. 1979 Oct 1;98(2):358–367. doi: 10.1016/0003-2697(79)90154-4. [DOI] [PubMed] [Google Scholar]
  15. Payne N. S., Sancar A. The LexA protein does not bind specifically to the two SOS box-like sequences immediately 5' to the phr gene. Mutat Res. 1989 Nov;218(3):207–210. doi: 10.1016/0921-8777(89)90005-0. [DOI] [PubMed] [Google Scholar]
  16. Perry K. L., Elledge S. J., Mitchell B. B., Marsh L., Walker G. C. umuDC and mucAB operons whose products are required for UV light- and chemical-induced mutagenesis: UmuD, MucA, and LexA proteins share homology. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4331–4335. doi: 10.1073/pnas.82.13.4331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rhoads D. B., Laimins L., Epstein W. Functional organization of the kdp genes of Escherichia coli K-12. J Bacteriol. 1978 Aug;135(2):445–452. doi: 10.1128/jb.135.2.445-452.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sadosky A. B., Davidson A., Lin R. J., Hill C. W. rhs gene family of Escherichia coli K-12. J Bacteriol. 1989 Feb;171(2):636–642. doi: 10.1128/jb.171.2.636-642.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sancar A., Rupert C. S. Cloning of the phr gene and amplification of photolyase in Escherichia coli. Gene. 1978 Dec;4(4):295–308. doi: 10.1016/0378-1119(78)90047-1. [DOI] [PubMed] [Google Scholar]
  20. Sancar G. B., Smith F. W., Lorence M. C., Rupert C. S., Sancar A. Sequences of the Escherichia coli photolyase gene and protein. J Biol Chem. 1984 May 10;259(9):6033–6038. [PubMed] [Google Scholar]
  21. Sancar G. B., Smith F. W., Sancar A. Identification and amplification of the E. coli phr gene product. Nucleic Acids Res. 1983 Oct 11;11(19):6667–6678. doi: 10.1093/nar/11.19.6667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES