Characterization of a dam mutant of Serratia marcescens and nucleotide sequence of the dam region

The DNA of Serratia marcescens has N6-adenine methylation in GATC sequences. Among 2-aminopurine-sensitive mutants isolated from S. marcescens Sr41, one was identified which lacked GATC methylation. The mutant showed up to 30-fold increased spontaneous mutability and enhanced mutability after treatm...

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Bibliographic Details
Published in:Journal of bacteriology 1999-07, Vol.181 (13), p.3880-3885
Main Authors: Ostendorf, T, Cherepanov, P, de Vries, J, Wackernagel, W
Format: Article
Language:English
Subjects:
Bacteria
Bacteriology
Cloning, Molecular
Deoxyribonucleic acid
DNA
DNA Repair
Enzymes
Escherichia coli - genetics
Escherichia coli Proteins
Genes
Genes, Bacterial
Genetic Complementation Test
Genetics
Genetics and Molecular Biology
Molecular Sequence Data
Mutagenesis - genetics
Mutation
Restriction Mapping
Sequence Analysis, DNA
Serratia marcescens
Serratia marcescens - enzymology
Serratia marcescens - genetics
Site-Specific DNA-Methyltransferase (Adenine-Specific) - genetics
Species Specificity
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Summary:The DNA of Serratia marcescens has N6-adenine methylation in GATC sequences. Among 2-aminopurine-sensitive mutants isolated from S. marcescens Sr41, one was identified which lacked GATC methylation. The mutant showed up to 30-fold increased spontaneous mutability and enhanced mutability after treatment with 2-aminopurine, ethyl methanesulfonate, or UV light. The gene (dam) coding for the adenine methyltransferase (Dam enzyme) of S. marcescens was identified on a gene bank plasmid which alleviated the 2-aminopurine sensitivity and the higher mutability of a dam-13::Tn9 mutant of Escherichia coli. Nucleotide sequencing revealed that the deduced amino acid sequence of Dam (270 amino acids; molecular mass, 31.3 kDa) has 72% identity to the Dam enzyme of E. coli. The dam gene is located between flanking genes which are similar to those found to the sides of the E. coli dam gene. The results of complementation studies indicated that like Dam of E. coli and unlike Dam of Vibrio cholerae, the Dam enzyme of S. marcescens plays an important role in mutation avoidance by allowing the mismatch repair enzymes to discriminate between the parental and newly synthesized strands during correction of replication errors.
ISSN:0021-9193
1098-5530
DOI:10.1128/jb.181.13.3880-3885.1999