• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Mar 1988; 85(6): 1816–1820.
PMCID: PMC279870

RecA-mediated cleavage activates UmuD for mutagenesis: mechanistic relationship between transcriptional derepression and posttranslational activation.


The products of the SOS-regulated umuDC operon are required for most UV and chemical mutagenesis in Escherichia coli. It has been shown that the UmuD protein shares homology with LexA, the repressor of the SOS genes. In this paper we describe a series of genetic experiments that indicate that the purpose of RecA-mediated cleavage of UmuD at its bond between Cys-24 and Gly-25 is to activate UmuD for its role in mutagenesis and that the COOH-terminal fragment of UmuD is necessary and sufficient for the role of UmuD in UV mutagenesis. Other genetic experiments are presented that (i) support the hypothesis that the primary role of Ser-60 in UmuD function is to act as a nucleophile in the RecA-mediated cleavage reaction and (ii) raise the possibility that RecA has a third role in UV mutagenesis besides mediating the cleavage of LexA and UmuD.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Elledge SJ, Walker GC. Proteins required for ultraviolet light and chemical mutagenesis. Identification of the products of the umuC locus of Escherichia coli. J Mol Biol. 1983 Feb 25;164(2):175–192. [PubMed]
  • Shinagawa H, Kato T, Ise T, Makino K, Nakata A. Cloning and characterization of the umu operon responsible for inducible mutagenesis in Escherichia coli. Gene. 1983 Aug;23(2):167–174. [PubMed]
  • Walker GC. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. [PMC free article] [PubMed]
  • Walker GC. Inducible DNA repair systems. Annu Rev Biochem. 1985;54:425–457. [PubMed]
  • Kato T, Shinoura Y. Isolation and characterization of mutants of Escherichia coli deficient in induction of mutations by ultraviolet light. Mol Gen Genet. 1977 Nov 14;156(2):121–131. [PubMed]
  • Steinborn G. Uvm mutants of Escherichia coli K12 deficient in UV mutagenesis. I. Isolation of uvm mutants and their phenotypical characterization in DNA repair and mutagenesis. Mol Gen Genet. 1978 Sep 20;165(1):87–93. [PubMed]
  • Perry KL, Elledge SJ, Mitchell BB, Marsh L, Walker GC. 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. [PMC free article] [PubMed]
  • Kitagawa Y, Akaboshi E, Shinagawa H, Horii T, Ogawa H, Kato T. Structural analysis of the umu operon required for inducible mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4336–4340. [PMC free article] [PubMed]
  • Perry KL, Walker GC. Identification of plasmid (pKM101)-coded proteins involved in mutagenesis and UV resistance. Nature. 1982 Nov 18;300(5889):278–281. [PubMed]
  • Bagg A, Kenyon CJ, Walker GC. Inducibility of a gene product required for UV and chemical mutagenesis in Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5749–5753. [PMC free article] [PubMed]
  • Witkin EM. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. [PMC free article] [PubMed]
  • Little JW, Mount DW. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. [PubMed]
  • Little JW, Edmiston SH, Pacelli LZ, Mount DW. Cleavage of the Escherichia coli lexA protein by the recA protease. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3225–3229. [PMC free article] [PubMed]
  • Little JW. Autodigestion of lexA and phage lambda repressors. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1375–1379. [PMC free article] [PubMed]
  • Slilaty SN, Little JW. Lysine-156 and serine-119 are required for LexA repressor cleavage: a possible mechanism. Proc Natl Acad Sci U S A. 1987 Jun;84(12):3987–3991. [PMC free article] [PubMed]
  • Sauer RT, Yocum RR, Doolittle RF, Lewis M, Pabo CO. Homology among DNA-binding proteins suggests use of a conserved super-secondary structure. Nature. 1982 Jul 29;298(5873):447–451. [PubMed]
  • Roberts JW, Roberts CW, Craig NL. Escherichia coli recA gene product inactivates phage lambda repressor. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4714–4718. [PMC free article] [PubMed]
  • Sauer RT, Ross MJ, Ptashne M. Cleavage of the lambda and P22 repressors by recA protein. J Biol Chem. 1982 Apr 25;257(8):4458–4462. [PubMed]
  • Marsh L, Walker GC. New phenotypes associated with mucAB: alteration of a MucA sequence homologous to the LexA cleavage site. J Bacteriol. 1987 May;169(5):1818–1823. [PMC free article] [PubMed]
  • Blanco M, Herrera G, Collado P, Rebollo JE, Botella LM. Influence of RecA protein on induced mutagenesis. Biochimie. 1982 Aug-Sep;64(8-9):633–636. [PubMed]
  • Ennis DG, Fisher B, Edmiston S, Mount DW. Dual role for Escherichia coli RecA protein in SOS mutagenesis. Proc Natl Acad Sci U S A. 1985 May;82(10):3325–3329. [PMC free article] [PubMed]
  • Tessman ES, Peterson PK. Isolation of protease-proficient, recombinase-deficient recA mutants of Escherichia coli K-12. J Bacteriol. 1985 Aug;163(2):688–695. [PMC free article] [PubMed]
  • Bridges BA, Woodgate R. Mutagenic repair in Escherichia coli: products of the recA gene and of the umuD and umuC genes act at different steps in UV-induced mutagenesis. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4193–4197. [PMC free article] [PubMed]
  • Tessman ES, Tessman I, Peterson PK, Forestal JD. Roles of RecA protease and recombinase activities of Escherichia coli in spontaneous and UV-induced mutagenesis and in Weigle repair. J Bacteriol. 1986 Dec;168(3):1159–1164. [PMC free article] [PubMed]
  • Witkin EM, Kogoma T. Involvement of the activated form of RecA protein in SOS mutagenesis and stable DNA replication in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7539–7543. [PMC free article] [PubMed]
  • Ruiz-Rubio M, Woodgate R, Bridges BA, Herrera G, Blanco M. New role for photoreversible pyrimidine dimers in induction of prototrophic mutations in excision-deficient Escherichia coli by UV light. J Bacteriol. 1986 Jun;166(3):1141–1143. [PMC free article] [PubMed]
  • Shinagawa H, Iwasaki H, Kato T, Nakata A. RecA protein-dependent cleavage of UmuD protein and SOS mutagenesis. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1806–1810. [PMC free article] [PubMed]
  • Burckhardt SE, Woodgate R, Scheuermann RH, Echols H. UmuD mutagenesis protein of Escherichia coli: overproduction, purification, and cleavage by RecA. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1811–1815. [PMC free article] [PubMed]
  • Zagursky RJ, Berman ML. Cloning vectors that yield high levels of single-stranded DNA for rapid DNA sequencing. Gene. 1984 Feb;27(2):183–191. [PubMed]
  • Krueger JH, Elledge SJ, Walker GC. Isolation and characterization of Tn5 insertion mutations in the lexA gene of Escherichia coli. J Bacteriol. 1983 Mar;153(3):1368–1378. [PMC free article] [PubMed]
  • Sancar A, Wharton RP, Seltzer S, Kacinski BM, Clarke ND, Rupp WD. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. [PubMed]
  • Gimble FS, Sauer RT. Lambda repressor inactivation: properties of purified ind- proteins in the autodigestion and RecA-mediated cleavage reactions. J Mol Biol. 1986 Nov 5;192(1):39–47. [PubMed]
  • Ben-Bassat A, Bauer K, Chang SY, Myambo K, Boosman A, Chang S. Processing of the initiation methionine from proteins: properties of the Escherichia coli methionine aminopeptidase and its gene structure. J Bacteriol. 1987 Feb;169(2):751–757. [PMC free article] [PubMed]
  • Roberts JW, Roberts CW. Two mutations that alter the regulatory activity of E. coli recA protein. Nature. 1981 Apr 2;290(5805):422–424. [PubMed]
  • Elledge SJ, Walker GC. The muc genes of pKM101 are induced by DNA damage. J Bacteriol. 1983 Sep;155(3):1306–1315. [PMC free article] [PubMed]
  • Devoret R, Pierre M, Moreau PL. Prophage phi 80 is induced in Escherichia coli K12 recA430. Mol Gen Genet. 1983;189(2):199–206. [PubMed]
  • Kawashima H, Horii T, Ogawa T, Ogawa H. Functional domains of Escherichia coli recA protein deduced from the mutational sites in the gene. Mol Gen Genet. 1984;193(2):288–292. [PubMed]
  • Walker GC, Marsh L, Dodson LA. Genetic analyses of DNA repair: inference and extrapolation. Annu Rev Genet. 1985;19:103–126. [PubMed]
  • Fersht AR, Knill-Jones JW. Contribution of 3' leads to 5' exonuclease activity of DNA polymerase III holoenzyme from Escherichia coli to specificity. J Mol Biol. 1983 Apr 25;165(4):669–682. [PubMed]
  • Lu C, Scheuermann RH, Echols H. Capacity of RecA protein to bind preferentially to UV lesions and inhibit the editing subunit (epsilon) of DNA polymerase III: a possible mechanism for SOS-induced targeted mutagenesis. Proc Natl Acad Sci U S A. 1986 Feb;83(3):619–623. [PMC free article] [PubMed]
  • Kruger K, Grabowski PJ, Zaug AJ, Sands J, Gottschling DE, Cech TR. Self-splicing RNA: autoexcision and autocyclization of the ribosomal RNA intervening sequence of Tetrahymena. Cell. 1982 Nov;31(1):147–157. [PubMed]
  • Hunkapiller MW, Smallcombe SH, Whitaker DR, Richards JH. Carbon nuclear magnetic resonance studies of the histidine residue in alpha-lytic protease. Implications for the catalytic mechanism of serine proteases. Biochemistry. 1973 Nov 6;12(23):4732–4743. [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...