Method for the Isolation of Escherichia Coli Mutants with Enhanced Recombination Between Chromosomal Duplications

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1977 Apr 1

PMID 323226

Citations 86

Authors

E B Konrad

Affiliations

Soon will be listed here.

Abstract

A method is described for the isolation of Escherichia coli mutants that show increased recombination between a pair of chromosomal duplications. These "hyper-rec" mutants display a variety of secondary phenotypes. I have isolated a large number of hyper-rec mutants and found them useful in screening for mutants that accumulate labeled DNA fragments after short pulses with [3H]thymidine. The mutants so recovered include ones that are defective in deoxyribonucleic acid ligase, deoxyribonucleic acid polymerase I and its associated 5' yields 3' exonuclease, and a group of mutants, dnaS, that accumulate abnormally short Okazaki fragments. Evidence is presented that suggests that the lac-att80 segment of the chromosome cannot be inverted.

Citing Articles

Modulation of RecFORQ- and RecA-Mediated Homologous Recombination in Escherichia coli by Isoforms of Translation Initiation Factor IF2.

Mallikarjun J, SaiSree L, Himabindu P, Anupama K, Reddy M, Gowrishankar J J Bacteriol. 2022; 204(4):e0056921.

PMID: 35343793 PMC: 9017302. DOI: 10.1128/jb.00569-21.

Single Strand Annealing Plays a Major Role in RecA-Independent Recombination between Repeated Sequences in the Radioresistant Deinococcus radiodurans Bacterium.

Ithurbide S, Bentchikou E, Coste G, Bost B, Servant P, Sommer S PLoS Genet. 2015; 11(10):e1005636.

PMID: 26517555 PMC: 4627823. DOI: 10.1371/journal.pgen.1005636.

DNA Mismatch Repair.

Marinus M EcoSal Plus. 2015; 5(1).

PMID: 26442827 PMC: 4231543. DOI: 10.1128/ecosalplus.7.2.5.

Homologous Recombination-Experimental Systems, Analysis, and Significance.

Kuzminov A EcoSal Plus. 2015; 4(2).

PMID: 26442506 PMC: 4190071. DOI: 10.1128/ecosalplus.7.2.6.

Escherichia coli radD (yejH) gene: a novel function involved in radiation resistance and double-strand break repair.

Chen S, Byrne R, Wood E, Cox M Mol Microbiol. 2014; 95(5):754-68.

PMID: 25425430 PMC: 4339530. DOI: 10.1111/mmi.12885.

References

Beckwith J . A DELETION ANALYSIS OF THE LAC OPERATOR REGION IN ESCHERICHIA COLI. J Mol Biol. 1964; 8:427-30. DOI: 10.1016/s0022-2836(64)80206-0. View

Jacob F, MONOD J . Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961; 3:318-56. DOI: 10.1016/s0022-2836(61)80072-7. View

Tye B, Nyman P, Lehman I, Hochhauser S, Weiss B . Transient accumulation of Okazaki fragments as a result of uracil incorporation into nascent DNA. Proc Natl Acad Sci U S A. 1977; 74(1):154-7. PMC: 393216. DOI: 10.1073/pnas.74.1.154. View

Okazaki R, Arisawa M, Sugino A . Slow joining of newly replicated DNA chains in DNA polymerase I-deficient Escherichia coli mutants. Proc Natl Acad Sci U S A. 1971; 68(12):2954-7. PMC: 389568. DOI: 10.1073/pnas.68.12.2954. View

Hirota Y, RYTER A, Jacob F . Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968; 33:677-93. DOI: 10.1101/sqb.1968.033.01.077. View

Gottesman M, Hicks M, Gellert M . Genetics and function of DNA ligase in Escherichia coli. J Mol Biol. 1973; 77(4):531-47. DOI: 10.1016/0022-2836(73)90221-0. View

Konrad E, Modrich P, Lehman I . Genetic and enzymatic characterization of a conditional lethal mutant of Escherichia coli K12 with a temperature-sensitive DNA ligase. J Mol Biol. 1973; 77(4):519-29. DOI: 10.1016/0022-2836(73)90220-9. View

Bachmann B, Low K, Taylor A . Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976; 40(1):116-67. PMC: 413944. DOI: 10.1128/br.40.1.116-167.1976. View

SIEGEL E . Ultraviolet-sensitive mutator strain of Escherichia coli K-12. J Bacteriol. 1973; 113(1):145-60. PMC: 251612. DOI: 10.1128/jb.113.1.145-160.1973. View

10.

Konrad E, Lehman I . Novel mutants of Escherichia coli that accumulate very small DNA replicative intermediates. Proc Natl Acad Sci U S A. 1975; 72(6):2150-4. PMC: 432714. DOI: 10.1073/pnas.72.6.2150. View

11.

Olivera R, Bonhoeffer E . Replication of Escherichia coli requires DNA polymerase I. Nature. 1974; 250(5466):513-4. DOI: 10.1038/250513a0. View

12.

Konrad E, Lehman I . A conditional lethal mutant of Escherichia coli K12 defective in the 5' leads to 3' exonuclease associated with DNA polymerase I. Proc Natl Acad Sci U S A. 1974; 71(5):2048-51. PMC: 388383. DOI: 10.1073/pnas.71.5.2048. View

13.

Marinus M, Morris N . Biological function for 6-methyladenine residues in the DNA of Escherichia coli K12. J Mol Biol. 1974; 85(2):309-22. DOI: 10.1016/0022-2836(74)90366-0. View

14.

Modrich P, Lehman I . Enzymatic joining of polynucleotides. IX. A simple and rapid assay of polynucleotide joining (ligase) activity by measurement of circle formation from linear deoxyadenylate-deoxythymidylate copolymer. J Biol Chem. 1970; 245(14):3626-31. View

15.

KUEMPEL P, Veomett G . A possible function of DNA polymerase in chromosome replication. Biochem Biophys Res Commun. 1970; 41(4):973-80. DOI: 10.1016/0006-291x(70)90180-4. View

16.

Pauling C, Hamm L . Properties of a temperature-sensitive, radiation-sensitive mutant of Escherichia coli. II. DNA replication. Proc Natl Acad Sci U S A. 1969; 64(4):1195-202. PMC: 223268. DOI: 10.1073/pnas.64.4.1195. View

17.

DE LUCIA P, Cairns J . Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969; 224(5225):1164-6. DOI: 10.1038/2241164a0. View

18.

Pauling C, Hamm L . Properties of a temperature-sensitive radiation-sensitive mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1968; 60(4):1495-502. PMC: 224946. DOI: 10.1073/pnas.60.4.1495. View