Abstract
The close linkage of the glnA gene with polA was exploited to construct a fine structure map of polA by means of generalized transduction with phage P1. Nine different polA- alleles were mapped by recombinational crosses. The results indicate a gene order consistent with previous observations (Kelley and Grindley 1976a; Murray and Kelley 1979). Three mutations, polA5, polA6 and polA12 map within the "carboxy-terminal" or "large-fragment" portion of the gene in unambiguous order. Four alleles, known to affect the "aminoterminal" portion of the gene, polA107, polA214, polA480ex and polA4113, appear to be closely linked with certain ambiguities in their exact order. All four of these mutations are known to alter the 5'→3' exonuclease activity of DNA polymerase I and three of them result in the conditional lethal polA- phenotype. The polA1 nonsense mutation maps between these two groups in a position consistent with its known effect, production of an amber fragment that includes the 5'→3' exonuclease. The final allele, resA1, is another nonsense mutation that maps at the extreme "amino-terminus" of the cistron.——A number of control experiments were conducted to determine the effects of polA- mutations on the P1-mediated recombinational event. These experiments indicated that abortive transduction occurs quite frequently, but the formation of abortive transductants and segregation of unselected transduced markers among daughter progeny is like that observed by other investigators. There was no evidence that any individual polA- allele behaved in an exceptional fashion during recombination.
Full Text
The Full Text of this article is available as a PDF (1.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bambara R. A., Uyemura D., Lehman I. R. On the processive mechanism of Escherichia coli DNA polymerase I. Delayed initiation of polymerization. J Biol Chem. 1976 Jul 10;251(13):4090–4094. [PubMed] [Google Scholar]
- Friedberg E. C., Lehman I. R. Excision of thymine dimers by proteolytic and amber fragments of E. coli DNA polymerase I. Biochem Biophys Res Commun. 1974 May 7;58(1):132–139. doi: 10.1016/0006-291x(74)90901-2. [DOI] [PubMed] [Google Scholar]
- Grindley N. D., Kelley W. S. Effects of different alleles of the E. coli K12 pol A gene on the replication of non-transferring plasmids. Mol Gen Genet. 1976 Feb 2;143(3):311–318. doi: 10.1007/BF00269409. [DOI] [PubMed] [Google Scholar]
- Gross J., Gross M. Genetic analysis of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1166–1168. doi: 10.1038/2241166a0. [DOI] [PubMed] [Google Scholar]
- Heijneker H. L., Ellens D. J., Tjeerde R. H., Glickman B. W., van Dorp B., Pouwels P. H. A mutant of Escherichia coli K12 deficient in the 5'-3' exonucleolytic activity of DNA polymerase I. II. Purification and properties of the mutant enzyme. Mol Gen Genet. 1973 Jul 31;124(1):83–96. doi: 10.1007/BF00267167. [DOI] [PubMed] [Google Scholar]
- Hours C., Denhardt D. T. "Nick translation" in Escherichia coli rep strains deficient in DNA polymerase I activities. Mol Gen Genet. 1979 Apr 17;172(1):73–80. doi: 10.1007/BF00276217. [DOI] [PubMed] [Google Scholar]
- Jacobsen H., Klenow H., Overgaard-Hansen K. The N-terminal amino-acid sequences of DNA polymerase I from Escherichia coli and of the large and the small fragments obtained by a limited proteolysis. Eur J Biochem. 1974 Jun 15;45(2):623–627. doi: 10.1111/j.1432-1033.1974.tb03588.x. [DOI] [PubMed] [Google Scholar]
- Kelley W. S., Whitfield H. J. Purification of an altered DNA polymerase from an E. coli strain with a pol mutation. Nature. 1971 Mar 5;230(5288):33–36. doi: 10.1038/230033a0. [DOI] [PubMed] [Google Scholar]
- Konrad E. B., Lehman I. R. 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 May;71(5):2048–2051. doi: 10.1073/pnas.71.5.2048. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lehman I. R., Chien J. R. Persistence of deoxyribonucleic acid polymerase I and its 5'--3' exonuclease activity in PolA mutants of Escherichia coli K12. J Biol Chem. 1973 Nov 25;248(22):7717–7723. [PubMed] [Google Scholar]
- Murray N. E., Kelley W. S. Characterization of lambdapolA transducing phages; effective expression of the E. coli polA gene. Mol Gen Genet. 1979 Aug;175(1):77–87. doi: 10.1007/BF00267858. [DOI] [PubMed] [Google Scholar]
- Olivera R. M., Bonhoeffer E. Replication of Escherichia coli requires DNA polymerase I. Nature. 1974 Aug 9;250(5466):513–514. doi: 10.1038/250513a0. [DOI] [PubMed] [Google Scholar]
- Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]