Mutator mutations in Escherichia coli induced by the insertionof phage Mu and the transposable resistance elements Tn5 and Tn10

doi:10.1016/0027-5107(82)90122-1

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis

Volume 93, Issue 1, March 1982, Pages 25-33

https://doi.org/10.1016/0027-5107(82)90122-1 Get rights and content

Abstract

Mutator mutations in the mutS gene induced by the insertion of phage Mu or the transposable resistance elements Tn5 or Tn10 and those in the mutL gene induced by Tn5 and T10 gave mutagenic activities similar to that of the previously described mutS3 and mutL25 mutations. Various combinations of mutS::Tn5,mutL::Tn5, uvrE156, and the deletion mutation δmutH2 did not produce an additive effect. This supports the idea that the products of these genes function in the same pathway of error correction during DNA synthesis.

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There are more references available in the full text version of this article.

Cited by (46)

Mutational consequences of dNTP pool imbalances in E. coli
2013, DNA Repair
Citation Excerpt :
Two mismatch-repair-deficient (mutL) derivatives of KA796 were used: NR9559 (mutL211∷Tn5) [27] and NR12443 (mutL218∷Tn10). The mutL markers in the two strains derive, respectively, from strains ES1301 and ES1481 [28]. The dcd-12∷Tn10dkan allele [15] was introduced into KA796 and NR9559 (mutL) from BW1040 [15] (provided by Dr. Bernard Weiss, University of Michigan) yielding, respectively, NR12572 (dcd) and NR12573 (mutL dcd).
The accuracy of DNA synthesis depends on the accuracy of the polymerase as well as the quality and concentration(s) of the available 5′-deoxynucleoside-triphosphate DNA precursors (dNTPs). The relationships between dNTPs and error rates have been studied in vitro, but only limited insights exist into these correlations during in vivo replication. We have investigated this issue in the bacterium Escherichia coli by analyzing the mutational properties of dcd and ndk strains. These strains, defective in dCTP deaminase and nucleoside diphosphate kinase, respectively, are characterized by both disturbances of dNTP pools and a mutator phenotype. ndk strains have been studied before, but were included in this study, as controversies exist regarding the source of its mutator phenotype. We show that dcd strains suffer from increased intracellular levels of dCTP (4-fold) and reduced levels of dGTP (2-fold), while displaying, as measured using a set of lacZ reversion markers in a mismatch-repair defective (mutL) background, a strong mutator effect for G·C→T·A and A·T→T·A transversions (27- and 42-fold enhancement, respectively). In contrast, ndk strains possess a lowered dATP level (4-fold) and modestly enhanced dCTP level (2-fold), while its mutator effect is specific for just the A·T→T·A transversions. The two strains also display differential mutability for rifampicin-resistant mutants. Overall, our analysis reveals for both strains a satisfactory correlation between dNTP pool alterations and the replication error rates, and also suggests that a minimal explanation for the ndk mutator does not require assumptions beyond the predicted effect of the dNTP pools.
Specialized mismatch repair function of Glu339 in the Phe-X-Glu motif of yeast Msh6
2007, DNA Repair
The major eukaryotic mismatch repair (MMR) pathway requires Msh2–Msh6, which, like Escherichia coli MutS, binds to and participates in repair of the two most common replication errors, single base–base and single base insertion–deletion mismatches. For both types of mismatches, the side chain of E. coli Glu38 in a conserved Phe-X-Glu motif interacts with a mismatched base. The Oɛ of Glu38 forms a hydrogen bond with either the N7 of purines or the N3 of pyrimidines. We show here that changing E. coli Glu38 to alanine results in nearly complete loss of repair of both single base–base and single base deletion mismatches. In contrast, a yeast strain with alanine replacing homologous Glu339 in Msh6 has nearly normal repair for insertion–deletion and most base–base mismatches, but is defective in repairing base–base mismatches characteristic of oxidative stress, e.g. 8-oxo-G·A mismatches. The results suggest that bacterial MutS and yeast Msh2–Msh6 differ in how they recognize and/or process replication errors involving undamaged bases, and that Glu339 in Msh6 may have a specialized role in repairing mismatches containing oxidized bases.
Cellular response to horizontally transferred DNA in Escherichia coli is tuned by DNA repair systems
2005, DNA Repair
We studied how DNA divergence between recombining DNAs and the mismatch repair system modulate the SOS response in Escherichia coli. The observed positive log-linear correlation between SOS induction and DNA divergence, and the negative correlation between SOS induction and frequency of recombination, suggest that the level of SOS induction precisely reflects the difficulty of RecA protein to initiate a productive strand exchange process. Our results suggest that the mismatch repair system could contribute to this SOS induction more by affecting the RecA-catalyzed homology search than by acting on mismatched recombination intermediates. The propensity of the recombination machinery to promote recombination between the blocks of sequences with the highest identity results in the increasing ratios of merodiploids (partial diploids) over genuine recombinants (homologous replacements) with increasing DNA divergence. We discuss the role of molecular mechanisms involved in the control of the recombination between diverged DNA sequences in the maintenance of genomic stability and genome evolution.
The putative anti-anti-sigma factor BldG is post-translationally modified by phosphorylation in Streptomyces coelicolor
2003, FEMS Microbiology Letters
The Streptomyces coelicolor bldG gene encodes a protein showing similarity to the SpoIIAA and RsbV anti-anti-sigma factors of Bacillus subtilis. Purified maltose binding protein-BldG could be phosphorylated in vitro by wild-type S. coelicolor crude extract, and both the phosphorylated and unphosphorylated forms of BldG could be detected in vivo using isoelectric focusing. ATP was shown to serve as the phosphoryl group donor, and phosphorylation of BldG was abolished when the putative phosphorylation site was changed from a serine to an alanine residue. A bldG mutant strain expressing the non-phosphorylatable BldG protein was unable to undergo morphological differentiation or produce antibiotics even after prolonged incubation, suggesting that phosphorylation of BldG is necessary for proper development in S. coelicolor.
Responses to the Major Acrolein-derived Deoxyguanosine Adduct in Escherichia coli
2001, Journal of Biological Chemistry
Acrolein, a reactive α,β-unsaturated aldehyde found ubiquitously in the environment and formed endogenously in mammalian cells, reacts with DNA to form an exocyclic DNA adduct, 3H-8-hydroxy-3-(β-d-2′-deoxyribofuranosyl)-5,6,7,8-tetrahydropyrido[3,2-a]purine-9-one (γ-OH-PdG). The cellular processing and mutagenic potential of γ-OH-PdG have been examined, using a site-specific approach in which a single adduct is embedded in double-strand plasmid DNA. Analysis of progeny plasmid reveals that this adduct is excised by nucleotide excision repair. The apparent level of inhibition of DNA synthesis is ∼70% in Escherichia coli ΔrecA, uvrA. The block to DNA synthesis can be overcome partially byrecA-dependent recombination repair. Targeted G → T transversions were observed at a frequency of 7 × 10⁻⁴/translesion synthesis. Inactivation ofpolB, dinB, and umuD,C genes coding for “SOS” DNA polymerases did not affect significantly the efficiency or fidelity of translesion synthesis. In vitroprimer extension experiments revealed that the Klenow fragment of polymerase I catalyzes error-prone synthesis, preferentially incorporating dAMP and dGMP opposite γ-OH-PdG. We conclude from this study that DNA polymerase III catalyzes translesion synthesis across γ-OH-PdG in an error-free manner. Nucleotide excision repair, recombination repair, and highly accurate translesion synthesis combine to protect E. coli from the potential genotoxicity of this DNA adduct.
DNA replication errors produced by the replicative apparatus of Escherichia coli
1999, Journal of Molecular Biology
It has been hard to detect forward mutations generated during DNA synthesis in vitro by replicative DNA polymerases, because of their extremely high fidelity and a high background level of pre-existing mutations in the single-stranded template DNA used. Using the oriC plasmid DNA replication in vitro system and the rpsL forward mutation assay, we examined the fidelity of DNA replication catalyzed by the replicative apparatus of Escherichia coli. Upon DNA synthesis by the fully reconstituted system, the frequency of rpsL⁻mutations in the product DNA was increased to 1.9×10⁻⁴, 50-fold higher than the background level of the template DNA. Among the mutations generated in vitro, single-base frameshifts predominated and occurred with a pattern similar to those induced in mismatch-repair deficient E. coli cells, indicating that the major replication error was slippage at runs of the same nucleotide. Large deletions and other structural alterations of DNA appeared to be induced also during the action of the replicative apparatus.

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Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis

Abstract

References (18)

Incidence of mutator strains in Escherichia coli and coliforms in nature

Mutation Res.

Translocatable elements in procaryotes

Cell

Linkage map of Escherichia coli K-12

Insertion and excision of the transposable kanamycin resistance determinant Tn5

Bacteriophage Mu-1-induced mutation to mutT in Escherichia coli

J. Bacteriol.

Bacterial mutator genes and control of spontaneous mutation

Annu. Rev. Genet.

Mutator gene studies in Escherichia coli: the mutS gene

Genetics

Insertion of the tetracycline resistance translocation unit Tn10 in the lac operon of Escherichia coli K12

Mol. Gen. Genet.

Escherichia coli mutator mutants deficient in methylation-instructed DNA mismatch correction

Cited by (46)

Mutational consequences of dNTP pool imbalances in E. coli

Specialized mismatch repair function of Glu339 in the Phe-X-Glu motif of yeast Msh6

Cellular response to horizontally transferred DNA in Escherichia coli is tuned by DNA repair systems

The putative anti-anti-sigma factor BldG is post-translationally modified by phosphorylation in Streptomyces coelicolor

Responses to the Major Acrolein-derived Deoxyguanosine Adduct in Escherichia coli

DNA replication errors produced by the replicative apparatus of Escherichia coli