Elevated expression of DNA polymerase II increases spontaneous mutagenesis in Escherichia coli

ABSTRACT The Escherichia coli β sliding clamp protein is proposed to play an important role in effecting switches between different DNA polymerases during replication, repair, and translesion DNA synthesis. We recently described how strains bearing the dnaN159 allele, which encodes a mutant form of the β clamp (β159), display a UV-sensitive phenotype that is suppressed by inactivation of DNA polymerase IV (M. D. Sutton, J. Bacteriol. 186:6738-6748, 2004). As part of an ongoing effort to understand mechanisms of DNA polymerase management in E. coli, we have further characterized effects of the dnaN159 allele on polymerase usage. Three of the five E.coli DNA polymerases (II, IV, and V) are regulated as part of the global SOS response. Our results indicate that elevated expression of the dinB-encoded polymerase IV is sufficient to result in conditional lethality of the dnaN159 strain. In contrast, chronically activated RecA protein, expressed from the recA730 allele, is lethal to the dnaN159 strain, and this lethality is suppressed by mutations that either mitigate RecA730 activity (i.e., ΔrecR), or impair the activities of DNA polymerase II or DNA polymerase V (i.e., ΔpolB or ΔumuDC). Thus, we have identified distinct genetic requirements whereby each of the three different SOS-regulated DNA polymerases are able to confer lethality upon the dnaN159 strain, suggesting the presence of multiple mechanisms by which the actions of the cell's different DNA polymerases are managed in vivo.

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DNA Polymerase II Supports the Replicative Bypass of N2-Alkyl-2′-deoxyguanosine Lesions in Escherichia coli Cells

Chemical Research in Toxicology ◽

10.1021/acs.chemrestox.0c00478 ◽

2021 ◽

Author(s):

Yinan Wang ◽

Jun Wu ◽

Jiabin Wu ◽

Yinsheng Wang

Keyword(s):

Escherichia Coli ◽

Dna Polymerase ◽

Escherichia Coli Cells ◽

Dna Polymerase Ii

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Escherichia coli DNA polymerase II is homologous to α-like DNA polymerases

MGG Molecular & General Genetics ◽

10.1007/bf00273583 ◽

1991 ◽

Vol 226-226 (1-2) ◽

pp. 24-33 ◽

Cited By ~ 26

Author(s):

Hiroshi Iwasaki ◽

Yoshizumi Ishino ◽

Hiroyuki Toh ◽

Atsuo Nakata ◽

Hideo Shinagawa

Keyword(s):

Escherichia Coli ◽

Dna Polymerase ◽

Dna Polymerases ◽

Dna Polymerase Ii

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A Novel DNA Polymerase Family Found inArchaea

Journal of Bacteriology ◽

10.1128/jb.180.8.2232-2236.1998 ◽

1998 ◽

Vol 180 (8) ◽

pp. 2232-2236 ◽

Cited By ~ 69

Author(s):

Yoshizumi Ishino ◽

Kayoko Komori ◽

Isaac K. O. Cann ◽

Yosuke Koga

Keyword(s):

Escherichia Coli ◽

Dna Polymerase ◽

Pyrococcus Furiosus ◽

Dna Polymerases ◽

Open Reading Frames ◽

Gene Products ◽

Polymerase Gene ◽

Hyperthermophilic Archaeon ◽

Dna Polymerase Ii ◽

Reading Frames

ABSTRACT One of the most puzzling results from the complete genome sequence of the methanogenic archaeon Methanococcus jannaschii was that the organism may have only one DNA polymerase gene. This is because no other DNA polymerase-like open reading frames (ORFs) were found besides one ORF having the typical α-like DNA polymerase (family B). Recently, we identified the genes of DNA polymerase II (the second DNA polymerase) from the hyperthermophilic archaeonPyrococcus furiosus, which has also at least one α-like DNA polymerase (T. Uemori, Y. Sato, I. Kato, H. Doi, and Y. Ishino, Genes Cells 2:499–512, 1997). The genes in M. jannaschiiencoding the proteins that are homologous to the DNA polymerase II ofP. furiosus have been located and cloned. The gene products of M. jannaschii expressed in Escherichia colihad both DNA polymerizing and 3′→5′ exonuclease activities. We propose here a novel DNA polymerase family which is entirely different from other hitherto-described DNA polymerases.

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