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. 1983 Jan;80(2):358–362. doi: 10.1073/pnas.80.2.358

Protein degradation in Escherichia coli: the lon gene controls the stability of sulA protein.

S Mizusawa, S Gottesman
PMCID: PMC393376  PMID: 6300834

Abstract

Escherichia coli lon mutants are defective in the ATP-dependent proteolysis of abnormal proteins. The mutants are also sensitive to ultraviolet light (UV) in that septation is inhibited after exposure to UV. sulA mutations, isolated as suppressors of UV sensitivity unlinked to lon, do not affect proteolysis but allow septation to occur after DNA damage. We have confirmed the hypothesis that the product of the sulA gene is degraded by lon proteolysis. If sulA (the product of sulA) is a UV-inducible division inhibitor, as suggested by a variety of experiments, lon (the product of lon) may regulate cell division by regulating the half-life of sulA. We cloned the sulA gene in a bacteriophage lambda vector from a plasmid carrying the ompA region of E. coli. An 18-kilodalton polypeptide was identified as the product of the sulA gene. Pulse-chase labeling demonstrated that the half-life of the sulA protein is 1.2 min in lon+ cells and 19 min in lon- cells. This work demonstrates that lon proteolysis affects the stability of a native E. coli protein.

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Selected References

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

  1. Beck E., Bremer E. Nucleotide sequence of the gene ompA coding the outer membrane protein II of Escherichia coli K-12. Nucleic Acids Res. 1980 Jul 11;8(13):3011–3027. doi: 10.1093/nar/8.13.3011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brent R., Ptashne M. Mechanism of action of the lexA gene product. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4204–4208. doi: 10.1073/pnas.78.7.4204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bukhari A. I., Zipser D. Mutants of Escherichia coli with a defect in the degradation of nonsense fragments. Nat New Biol. 1973 Jun 20;243(129):238–241. doi: 10.1038/newbio243238a0. [DOI] [PubMed] [Google Scholar]
  4. Charette M. F., Henderson G. W., Markovitz A. ATP hydrolysis-dependent protease activity of the lon (capR) protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4728–4732. doi: 10.1073/pnas.78.8.4728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chung C. H., Goldberg A. L. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931–4935. doi: 10.1073/pnas.78.8.4931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clewell D. B., Helinski D. R. Properties of a supercoiled deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry. 1970 Oct 27;9(22):4428–4440. doi: 10.1021/bi00824a026. [DOI] [PubMed] [Google Scholar]
  7. Enquist L. W., Weisberg R. A. The red plaque test: a rapid method for identification of excision defective variants of bacteriophage lambda. Virology. 1976 Jul 1;72(1):147–153. doi: 10.1016/0042-6822(76)90319-6. [DOI] [PubMed] [Google Scholar]
  8. Enquist L., Sternberg N. In vitro packaging of lambda Dam vectors and their use in cloning DNA fragments. Methods Enzymol. 1979;68:281–298. doi: 10.1016/0076-6879(79)68020-5. [DOI] [PubMed] [Google Scholar]
  9. Gayda R. C., Avni H., Berg P. E., Markovitz A. Outer membrane protein a and other polypeptides regulate capsular polysaccharide synthesis in E. coli K-12. Mol Gen Genet. 1979 Oct 1;175(3):325–332. doi: 10.1007/BF00397232. [DOI] [PubMed] [Google Scholar]
  10. Gayda R. C., Yamamoto L. T., Markovitz A. Second-site mutations in capR (lon) strains of Escherichia coli K-12 that prevent radiation sensitivity and allow bacteriophage lambda to lysogenize. J Bacteriol. 1976 Sep;127(3):1208–1216. doi: 10.1128/jb.127.3.1208-1216.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. George J., Castellazzi M., Buttin G. Prophage induction and cell division in E. coli. III. Mutations sfiA and sfiB restore division in tif and lon strains and permit the expression of mutator properties of tif. Mol Gen Genet. 1975 Oct 22;140(4):309–332. [PubMed] [Google Scholar]
  12. Gottesman M. E., Yarmolinsky M. B. Integration-negative mutants of bacteriophage lambda. J Mol Biol. 1968 Feb 14;31(3):487–505. doi: 10.1016/0022-2836(68)90423-3. [DOI] [PubMed] [Google Scholar]
  13. Gottesman S., Gottesman M., Shaw J. E., Pearson M. L. Protein degradation in E. coli: the lon mutation and bacteriophage lambda N and cII protein stability. Cell. 1981 Apr;24(1):225–233. doi: 10.1016/0092-8674(81)90518-3. [DOI] [PubMed] [Google Scholar]
  14. Gottesman S., Halpern E., Trisler P. Role of sulA and sulB in filamentation by lon mutants of Escherichia coli K-12. J Bacteriol. 1981 Oct;148(1):265–273. doi: 10.1128/jb.148.1.265-273.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gottesman S., Zipser D. Deg phenotype of Escherichia coli lon mutants. J Bacteriol. 1978 Feb;133(2):844–851. doi: 10.1128/jb.133.2.844-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HOWARD-FLANDERS P., SIMSON E., THERIOT L. A LOCUS THAT CONTROLS FILAMENT FORMATION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI K-12. Genetics. 1964 Feb;49:237–246. doi: 10.1093/genetics/49.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Henning U., Royer H. D., Teather R. M., Hindennach I., Hollenberg C. P. Cloning of the structural gene (ompA) for an integral outer membrane protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4360–4364. doi: 10.1073/pnas.76.9.4360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Herskowitz I., Hagen D. The lysis-lysogeny decision of phage lambda: explicit programming and responsiveness. Annu Rev Genet. 1980;14:399–445. doi: 10.1146/annurev.ge.14.120180.002151. [DOI] [PubMed] [Google Scholar]
  19. Huisman O., D'Ari R. An inducible DNA replication-cell division coupling mechanism in E. coli. Nature. 1981 Apr 30;290(5809):797–799. doi: 10.1038/290797a0. [DOI] [PubMed] [Google Scholar]
  20. Huisman O., D'Ari R., George J. Further characterization of sfiA and sfiB mutations in Escherichia coli. J Bacteriol. 1980 Oct;144(1):185–191. doi: 10.1128/jb.144.1.185-191.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jaskunas S. R., Lindahl L., Nomura M. Identification of two copies of the gene for the elongation factor EF-Tu in E. coli. Nature. 1975 Oct 9;257(5526):458–462. doi: 10.1038/257458a0. [DOI] [PubMed] [Google Scholar]
  22. Johnson B. F. Fine structure mapping and properties of mutations suppressing the lon mutation in Escherichia coli K-12 and B strains. Genet Res. 1977 Dec;30(3):273–286. doi: 10.1017/s0016672300017687. [DOI] [PubMed] [Google Scholar]
  23. Johnson B. F., Greenberg J. Mapping of sul, the suppressor of lon in Escherichia coli. J Bacteriol. 1975 May;122(2):570–574. doi: 10.1128/jb.122.2.570-574.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kenyon C. J., Walker G. C. DNA-damaging agents stimulate gene expression at specific loci in Escherichia coli. Proc Natl Acad Sci U S A. 1980 May;77(5):2819–2823. doi: 10.1073/pnas.77.5.2819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. King J., Laemmli U. K. Polypeptides of the tail fibres of bacteriophage T4. J Mol Biol. 1971 Dec 28;62(3):465–477. doi: 10.1016/0022-2836(71)90148-3. [DOI] [PubMed] [Google Scholar]
  26. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  27. Little J. W., Mount D. W., Yanisch-Perron C. R. Purified lexA protein is a repressor of the recA and lexA genes. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4199–4203. doi: 10.1073/pnas.78.7.4199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Movva N. R., Nakamura K., Inouye M. Amino acid sequence of the signal peptide of ompA protein, a major outer membrane protein of Escherichia coli. J Biol Chem. 1980 Jan 10;255(1):27–29. [PubMed] [Google Scholar]
  30. Sancar G. B., Sancar A., Little J. W., Rupp W. D. The uvrB gene of Escherichia coli has both lexA-repressed and lexA-independent promoters. Cell. 1982 Mar;28(3):523–530. doi: 10.1016/0092-8674(82)90207-0. [DOI] [PubMed] [Google Scholar]
  31. Shimada K., Campbell A. Int-constitutive mutants of bacteriophage lambda. Proc Natl Acad Sci U S A. 1974 Jan;71(1):237–241. doi: 10.1073/pnas.71.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shineberg B., Zipser D. The ion gene and degradation of beta-galactosidase nonsense fragments. J Bacteriol. 1973 Dec;116(3):1469–1471. doi: 10.1128/jb.116.3.1469-1471.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Simon L. D., Gottesman M., Tomczak K., Gottesman S. Hyperdegradation of proteins in Escherichia coli rho mutants. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1623–1627. doi: 10.1073/pnas.76.4.1623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Takano T. Bacterial mutants defective in plasmid formation: requirement for the lon + allele. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1469–1473. doi: 10.1073/pnas.68.7.1469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Van Alphen L., Havekes L., Lugtenberg B. Major outer membrane protein d of Escherichia coli K12. Purification and in vitro activity of bacteriophages k3 and f-pilus mediated conjugation. FEBS Lett. 1977 Mar 15;75(1):285–290. doi: 10.1016/0014-5793(77)80104-x. [DOI] [PubMed] [Google Scholar]
  36. Walker J. R., Ussery C. L., Allen J. S. Bacterial cell division regulation: lysogenization of conditional cell division lon - mutants of Escherichia coli by bacteriophage. J Bacteriol. 1973 Mar;113(3):1326–1332. doi: 10.1128/jb.113.3.1326-1332.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Witkin E. M. The radiation sensitivity of Escherichia coli B: a hypothesis relating filament formation and prophage induction. Proc Natl Acad Sci U S A. 1967 May;57(5):1275–1279. doi: 10.1073/pnas.57.5.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]

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