The Biochemical Basis and In Vivo Regulation of SOS-induced Mutagenesis Promoted by Escherichia coli DNA Polymerase V (UmuD'2C)

2000 ◽  
Vol 65 (0) ◽  
pp. 31-40 ◽  
Author(s):  
M.F. GOODMAN ◽  
R. WOODGATE
Keyword(s):  
Escherichia Coli ◽  
Dna Polymerase ◽  
Biochemical Basis ◽  
Dna Polymerase V ◽  
Induced Mutagenesis

scholarly journals Escherichia coli DNA Polymerase III Can Replicate Efficiently past a T-T cis-syn Cyclobutane Dimer if DNA Polymerase V and the 3′ to 5′ Exonuclease Proofreading Function Encoded by dnaQ Are Inactivated

2002 ◽  
Vol 184 (10) ◽  
pp. 2674-2681 ◽  
Author(s):  
Angela Borden ◽  
Paul I. O'Grady ◽  
Dominique Vandewiele ◽  
Antonio R. Fernández de Henestrosa ◽  
Christopher W. Lawrence ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Dna Polymerase ◽  
Wild Type ◽  
Lesion Bypass ◽  
Base Pairs ◽  
Pol V ◽  
Pol I ◽  
Cyclobutane Dimer ◽  
Dna Polymerase V ◽  
Pol Iii

ABSTRACT Although very little replication past a T-T cis-syn cyclobutane dimer normally takes place in Escherichia coli in the absence of DNA polymerase V (Pol V), we previously observed as much as half of the wild-type bypass frequency in Pol V-deficient (ΔumuDC) strains if the 3′ to 5′ exonuclease proofreading activity of the Pol III ε subunit was also disabled by mutD5. This observation might be explained in at least two ways. In the absence of Pol V, wild-type Pol III might bind preferentially to the blocked primer terminus but be incapable of bypass, whereas the proofreading-deficient enzyme might dissociate more readily, providing access to bypass polymerases. Alternatively, even though wild-type Pol III is generally regarded as being incapable of lesion bypass, proofreading-impaired Pol III might itself perform this function. We have investigated this issue by examining dimer bypass frequencies in ΔumuDC mutD5 strains that were also deficient for Pol I, Pol II, and Pol IV, both singly and in all combinations. Dimer bypass frequencies were not decreased in any of these strains and indeed in some were increased to levels approaching those found in strains containing Pol V. Efficient dimer bypass was, however, entirely dependent on the proofreading deficiency imparted by mutD5, indicating the surprising conclusion that bypass was probably performed by the mutD5 Pol III enzyme itself. This mutant polymerase does not replicate past the much more distorted T-T (6-4) photoadduct, however, suggesting that it may only replicate past lesions, like the T-T dimer, that form base pairs normally.

scholarly journals Inactivation of Umuc Protein Significantly Reduces Resistance to Ciprofloxacin and SOS Mutagenesis in Escherichia coli Mutants Harboring Intact Umud Gene

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Razieh Pourahmad Jaktaji ◽  
Sayedeh Marzieh Nourbakhsh Rezaei
Keyword(s):  
Escherichia Coli ◽  
Dna Polymerase ◽  
Genetic Background ◽  
Pcr Amplification ◽  
Sos Response ◽  
Rpob Gene ◽  
Dilution Method ◽  
E Coli ◽  
Sos Mutagenesis ◽  
Dna Polymerase V

Background: Ciprofloxacin induces SOS response and mutagenesis by activation of UmuD’2C (DNA polymerase V) and DinB (DNA polymerase IV) in Escherichia coli, leading to antibiotic resistance during therapy. Inactivation of DNA polymerase V can result in the inhibition of mutagenesis in E. coli. Objectives: The aim of this research was to investigate the effect of UmuC inactivation on resistance to ciprofloxacin and SOS mutagenesis in E. coli mutants. Methods: Ciprofloxacin-resistant mutants were produced in a umuC- genetic background in the presence of increasing concentrations of ciprofloxacin. The minimum inhibitory concentration of umuC-mutants was measured by broth dilution method. Alterations in the rifampin resistance-determing region of rpoB gene were assessed by PCR amplification and DNA sequencing. The expression of SOS genes was measured by quantitative real-time PCR assay. Results: Results showed that despite the induction of SOS response (overexpression of recA, dinB, and umuD genes) following exposure to ciprofloxacin in E. coliumuC mutants, resistance to ciprofloxacin and SOS mutagenesis significantly decreased. However, rifampicin-resistant clones emerged in this genetic background. One of these clones showed mutations in the rifampicin resistance-determining region of rpoB (cluster II). The low mutation frequency of E. coli might be associated with the presence and overexpression of umuD gene, which could somehow limit the activity of DinB, the location and type of mutations in the β subunit of RNA polymerase. Conclusions: In conclusion, for increasing the efficiency of ciprofloxacin against Gram-negative bacteria, use of an inhibitor of umuC, along with ciprofloxacin, would be helpful.

scholarly journals Escherichia coli DNA polymerase II catalyzes chromosomal and episomal DNA synthesis in vivo

1997 ◽  
Vol 94 (3) ◽  
pp. 946-951 ◽  
Author(s):  
S. Rangarajan ◽  
G. Gudmundsson ◽  
Z. Qiu ◽  
P. L. Foster ◽  
M. F. Goodman
Keyword(s):  
Escherichia Coli ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Dna Polymerase Ii ◽  
Episomal Dna

Viroid RNA is accepted as a template for in vitro transcription by DNA-dependent DNA polymerase I and RNA polymerase from Escherichia coli

1982 ◽  
Vol 2 (11) ◽  
pp. 929-939 ◽  
Author(s):  
Wolfgang Rohde ◽  
Hans-Richard Rackwitz ◽  
Frank Boege ◽  
Heinz L. Sänger
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Dna Polymerase ◽  
Unit Length ◽  
Dna Polymerase I ◽  
Experimental Conditions ◽  
In Vitro Synthesis ◽  
Polymerase I

The RNA genome of potato spindle tuber viroid (PSTV) is transcribed in vitro into complementary DNA and RNA by DNA-dependent DNA polymerase I and RNA polymerase, respectively, from Escherichia coli. In vitro synthesis of complementary RNA produces distinct transcripts larger than unit length thus reflecting the in vivo mechanism of viroid replication. The influence of varying experimental conditions on the transcription process is studied; actinomycin D is found to drastically reduce complementary RNA synthesis from the PSTV RNA template by RNA polymerase.

scholarly journals Role of Escherichia coli DNA Polymerase IV in In Vivo Replication Fidelity

2004 ◽  
Vol 186 (14) ◽  
pp. 4802-4807 ◽  
Author(s):  
Wojciech Kuban ◽  
Piotr Jonczyk ◽  
Damian Gawel ◽  
Karolina Malanowska ◽  
Roel M. Schaaper ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mismatch Repair ◽  
Dna Polymerase ◽  
Error Rate ◽  
The Other ◽  
Chromosomal Dna ◽  
Pol Iv ◽  
Dna Polymerase Iv

ABSTRACT We have investigated whether DNA polymerase IV (Pol IV; the dinB gene product) contributes to the error rate of chromosomal DNA replication in Escherichia coli. We compared mutation frequencies in mismatch repair-defective strains that were either dinB positive or dinB deficient, using a series of mutational markers, including lac targets in both orientations on the chromosome. Virtually no contribution of Pol IV to the chromosomal mutation rate was observed. On the other hand, a significant effect of dinB was observed for reversion of a lac allele when the lac gene resided on an F′(pro-lac) episome.

scholarly journals DNA Polymerase V Kinetics Support the Instructive Nature of an Oxidized Abasic Lesion in Escherichia coli

Biochemistry ◽  
2013 ◽  
Vol 52 (37) ◽  
pp. 6301-6303 ◽  
Author(s):  
John Ernest V. Bajacan ◽  
Marc M. Greenberg
Keyword(s):  
Escherichia Coli ◽  
Dna Polymerase ◽  
Dna Polymerase V
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