Multi-stage proofreading in DNA replication

1993 ◽  
Vol 26 (3) ◽  
pp. 225-331 ◽  
Author(s):  
Robert A. Beckman ◽  
Lawrence A. Loeb
Keyword(s):  
Steady State ◽  
Dna Replication ◽  
Dna Polymerase ◽  
Dna Polymerases ◽  
Kinetic Studies ◽  
Dna Polymerase I ◽  
E Coli ◽  
Multi Stage ◽  
Base Selection ◽  
Polymerase I

The mechanisms by which DNA polymerases achieve their remarkable fidelity, including base selection and proofreading, are briefly reviewed. Nine proofreading models from the current literature are evaluated in the light of steady-state and transient kinetic studies of E. coli DNA polymerase I, the beststudied DNA polymerase.

E. coli DNA Polymerase I and other Host Functions participate in T4 DNA Replication and Recombination

1972 ◽  
Vol 240 (96) ◽  
pp. 12-16 ◽  
Author(s):  
GISELA MOSIG ◽  
DONALD W. BOWDEN ◽  
SUSAN BOCK
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Dna Polymerase I ◽  
E Coli ◽  
Polymerase I

scholarly journals Nucleotide probes of DNA polymerases.

1996 ◽  
Vol 43 (1) ◽  
pp. 115-124 ◽  
Author(s):  
G E Wright
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerases ◽  
Site Directed Mutagenesis ◽  
Dna Polymerase I ◽  
E Coli ◽  
Enzyme Binding ◽  
The Family ◽  
Synthetic Approaches ◽  
Polymerase I ◽  
Related Compounds

The modified nucleotides, N2-(p-n-butylphenyl)dGTP and 2-(p-n-butylanilino) dATP and related compounds have been developed as inhibitor-probes of B family DNA polymerases. Synthetic approaches to these compounds are summarized. The nucleotides are potent, non-substrate inhibitors of DNA polymerase a. In contrast, they inhibit other members of the family with less potency but act as substrates for these enzymes. Modelling of the inhibitor: enzyme binding mechanism has been done based on the known structure of E. coli DNA polymerase I, and site-directed mutagenesis experiments to evaluate this mechanism are proposed.

A Sensitive Method for Identification of DNA Dependent DNA Polymerases in Acrylamide Gels after Seperation by Micro Disc Electrophoresis

1973 ◽  
Vol 28 (7-8) ◽  
pp. 376-378 ◽  
Author(s):  
E. Jürgen Zöllner ◽  
Werner E.G. Müller ◽  
Rudolf K. Zahn
Keyword(s):  
Dna Polymerase ◽  
Enzyme Assay ◽  
Dna Polymerases ◽  
Disc Electrophoresis ◽  
Dna Polymerase I ◽  
E Coli ◽  
Insoluble Polymer ◽  
Polymerase I ◽  
Sensitive Method ◽  
Bromide Complex

Abstract DNA polymerase, disc electrophoresis, template affinity Two sensitive methods are described for detection of DNA dependent DNA polymerase activities in polyacrylamide gels after their fractionation by micro-disc electrophoresis. One technique is based on the increase in fluorescence of the ethidium bromide complex with template polydeoxyribonucleotides brought about by the action of the polymerases. The sensitivity of the previously described technique has been enhanced. Another method, 14 fold as sensitive, uses radioactive precursors in the enzyme assay after electrophoretic separation; washing, slicing and counting allows to evaluate incorporation into acid insoluble polymer, requiring 3* 10" 2 units corresponding to approx. 0.02 /μg of E. coli DNA polymerase I preparation. Differing activities of polymerase species and differing template preferences may be investigated with these techniques.

Site-specific modification of E. coli DNA polymerase I large fragment with pyridoxal 5'-phosphate

Biochemistry ◽  
1984 ◽  
Vol 23 (9) ◽  
pp. 2073-2078 ◽  
Author(s):  
Anup K. Hazra ◽  
Sevilla Detera-Wadleigh ◽  
Samuel H. Wilson
Keyword(s):  
Dna Polymerase ◽  
Large Fragment ◽  
Dna Polymerase I ◽  
Site Specific ◽  
E Coli ◽  
Specific Modification ◽  
Polymerase I

scholarly journals Genetic evidence for two protein domains and a potential new activity in bacteriophage T4 DNA polymerase.

Genetics ◽  
1990 ◽  
Vol 124 (2) ◽  
pp. 213-220 ◽  
Author(s):  
L J Reha-Krantz
Keyword(s):  
Dna Polymerase ◽  
Bacteriophage T4 ◽  
Rnase H ◽  
Ribonuclease H ◽  
Temperature Sensitive ◽  
Polymerase Gene ◽  
Dna Polymerase I ◽  
E Coli ◽  
Intragenic Complementation ◽  
Polymerase I

Abstract Intragenic complementation was detected within the bacteriophage T4 DNA polymerase gene. Complementation was observed between specific amino (N)-terminal, temperature-sensitive (ts) mutator mutants and more carboxy (C)-terminal mutants lacking DNA polymerase polymerizing functions. Protein sequences surrounding N-terminal mutation sites are similar to sequences found in Escherichia coli ribonuclease H (RNase H) and in the 5'----3' exonuclease domain of E. coli DNA polymerase I. These observations suggest that T4 DNA polymerase, like E. coli DNA polymerase I, contains a discrete N-terminal domain.

scholarly journals The mechanism of recA polA lethality: suppression by RecA-independent recombination repair activated by the lexA(Def) mutation in Escherichia coli.

Genetics ◽  
1995 ◽  
Vol 139 (4) ◽  
pp. 1483-1494 ◽  
Author(s):  
Y Cao ◽  
T Kogoma
Keyword(s):  
Escherichia Coli ◽  
Dna Replication ◽  
Dna Polymerase ◽  
Reca Protein ◽  
Minor Role ◽  
Temperature Sensitive ◽  
Restrictive Temperature ◽  
Dna Polymerase I ◽  
Recombination Repair ◽  
Polymerase I

Abstract The mechanism of recA polA lethality in Escherichia coli has been studied. Complementation tests have indicated that both the 5'-->3' exonuclease and the polymerization activities of DNA polymerase I are essential for viability in the absence of RecA protein, whereas the viability and DNA replication of DNA polymerase I-defective cells depend on the recombinase activity of RecA. An alkaline sucrose gradient sedimentation analysis has indicated that RecA has only a minor role in Okazaki fragment processing. Double-strand break repair is proposed for the major role of RecA in the absence of DNA polymerase I. The lexA(Def)::Tn5 mutation has previously been shown to suppress the temperature-sensitive growth of recA200(Ts) polA25::spc mutants. The lexA(Def) mutation can alleviate impaired DNA synthesis in the recA200(Ts) polA25::spc mutant cells at the restrictive temperature. recF+ is essential for this suppression pathway. recJ and recQ mutations have minor but significant adverse effects on the suppression. The recA200(Ts) allele in the recA200(Ts) polA25::spc lexA(Def) mutant can be replaced by delta recA, indicating that the lexA(Def)-induced suppression is RecA independent. lexA(Def) reduces the sensitivity of delta recA polA25::spc cells to UV damage by approximately 10(4)-fold. lexA(Def) also restores P1 transduction proficiency to the delta recA polA25::spc mutant to a level that is 7.3% of the recA+ wild type. These results suggest that lexA(Def) activates a RecA-independent, RecF-dependent recombination repair pathway that suppresses the defect in DNA replication in recA polA double mutants.

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