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S M Picksley

Publications and source records attributed to S M Picksley.

23 records · Page 2Linked to original sources
PubMed1987

Nucleotide sequence and LexA regulation of the Escherichia coli recN gene.

The nucleotide sequence of a 2224 bp region of the Escherichia coli chromosome that carries the LexA regulated recN gene has been determined. A region of 1701 nucleotides encoding a polypeptide of 567 amino acids with a predicted molecular weight of 63,599 was identified as the most probable sequence for the recN structural gene. The proposed initiation codon is preceded by a reasonable Shine-Dalgarno sequence and a promoter region containing two 16 bp sequences, separated by 6 bp, that match the consensus sequence (SOS box) for binding LexA protein. DNA fragments containing this putative promoter region are shown to bind LexA in vitro and to have LexA-regulated promoter activity in vivo. The amino acid sequence of RecN predicted from the DNA contains a region that is homologous to highly conserved sequences found in several DNA repair enzymes and other proteins that bind ATP. A sequence of 9 amino acids was found to be homologous to a region of the RecA protein of E. coli postulated to have a role in DNA/nucleotide binding.

Amino Acid Sequence↗
PubMed1985

The recN locus of Escherichia coli K12: molecular analysis and identification of the gene product.

The recN gene which is necessary for inducible DNA repair and recombination in Escherichia coli has been cloned into the low copy plasmid vector pHSG415. Analysis of the recombinant plasmid, pSP100, revealed a 5.6 Kb HindIII insert of chromosomal DNA. Transposon inactivation of recN function and analysis of a recN::Mu(Ap lac) fusion located the coding region to a 1.4 Kb region within a 2.1 Kb BglII-AvaI DNA fragment transcribed in a clockwise direction with respect to the chromosome map. The gene product was identified in maxicells as a 60,000 dalton protein. Synthesis of this protein was increased in cells lacking LexA activity or in strains carrying recN cloned into the multicopy vector pBR322. Multiple copies of recN increase resistance to ionizing radiation in recN mutants but reduce the survival of a wild-type strain.

Bacterial Proteins↗
PubMed1984

Repair of DNA double-strand breaks in Escherichia coli K12 requires a functional recN product.

Mutation of the recN gene of Escherichia coli in a recBC sbcB genetic background blocks conjugational recombination and confers increased sensitivity to UV light and mitomycin C. The basis for this phenotype was investigated by monitoring the properties associated with recN mutations in otherwise wild-type strains. It was established that recN single mutants are almost fully resistant to UV irradiation, and that there is no detectable defect in repair of UV lesions by excision, error-prone, or recombinational mechanisms. However, recN mutations confer sensitivity to mitomycin C and ionizing radiation both in wild-type and recB sbcB strains. The sensitivity to ionizing radiation is correlated with a deficiency in the capacity to repair DNA double-strand breaks by a UV inducible mechanism. Recombinant lambda phages that complement the recombination and repair defects of recN recBC sbcB mutants have been identified, and the recN gene has been cloned from these phages into a low copy-number plasmid.

Bacterial Proteins↗
PubMed1983

Inducible expression of a gene specific to the RecF pathway for recombination in Escherichia coli K12.

The Mud(Aplac) operon fusion technique of Casadaban and Cohen (1979) was used to search for inducible functions specific to the RecF pathway of conjugal recombination. A fusion mutant of a recBC sbcB mutant which showed less than 1% of the normal level of recombination in Hfr crosses has been isolated and designated as rec-259. The mutation is shown to be closely linked to tyrA at approximately 57.5 min in relation to the standard genetic map, and is quite distinct from recA. Two point mutations within this gene have also been obtained. Mutation of this gene interferes specifically with the RecF pathway of recombination, and also causes increased sensitivity to mitomycin C and UV light. Expression of the lac genes in the rec-259 fusion strain is increased following damage to DNA, but not in lexA and recA derivatives. These observations demonstrate the existence of an inducible gene which is regulated by lexA and whose expression is required for RecF recombination and DNA repair.

DNA Repair↗