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2 edition of Deoxyribonucleotides as determinants of DNA replication fidelity in bacteriophage T4 found in the catalog.

Deoxyribonucleotides as determinants of DNA replication fidelity in bacteriophage T4

Roy Geoffrey Sargent

Deoxyribonucleotides as determinants of DNA replication fidelity in bacteriophage T4

by Roy Geoffrey Sargent

  • 298 Want to read
  • 24 Currently reading

Published .
Written in English

    Subjects:
  • DNA -- Synthesis.,
  • Bacteriophage T4.

  • Edition Notes

    Statementby Roy Geoffrey Sargent.
    The Physical Object
    Pagination205 leaves, bound :
    Number of Pages205
    ID Numbers
    Open LibraryOL14280337M

    dna replication. Web. Medical Information Search including key DNA replication factors and proteins associated with transcription, chromatin organization, DNA repair and mRNA A global analysis of DNA replication initiation in T. brucei showed that TbORC1 (subunit of the origin recognition complex, ORC Tb proved to be essential since its silencing caused a growth defect.   Increased fidelity can also be obtained if DNA polymerases check the accuracy of the bound nucleotide in more than one step (11, 12).Selection may be exerted in an initial binding step to form pre-insertion complexes, in a post-binding step that evaluates the accuracy of the newly forming base pair by an induced-fit mechanism (reviewed by Johnson, 13), and in the chemical step of.

    De Waard A, Paul AV, Lehman IR. The structural gene for deoxyribonucleic acid polymerase in bacteriophages T4 and T5. Proc Natl Acad Sci U S A. Oct; 54 (4)– [PMC free article] Fersht AR. Fidelity of replication of phage phi X DNA by DNA polymerase III holoenzyme: spontaneous mutation by misincorporation. deoxyribonucleic acid (DNA) precursors and the replication of DNA. The faithful replication of DNA is critical to the preservation of species; on the other hand, mutation contributes to evolution. Among the determinants of replication fidelity are the relative concentrations of the deoxyribonucleotide precursors of DNA (Weymouth et al.

    The Proofreading Pathway of Bacteriophage T4 DNA Polymerase. Journal of Biological Chemistry , (36), DOI: /jbc Lisa C Kroutil, Michelle West Frey, Barbara F Kaboord, Thomas A Kunkel, Stephen J Benkovic. Effect of accessory proteins on T4 DNA polymerase replication fidelity. For > 35 yr, we have known that the accuracy of DNA replication is controlled in large part by the relative concentrations of the 4 canonical deoxyribonucleoside 5'‐triphosphates (dNTPs) at the rep.


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Deoxyribonucleotides as determinants of DNA replication fidelity in bacteriophage T4 by Roy Geoffrey Sargent Download PDF EPUB FB2

I describe in this dissertation the further development of a procaryotic system, namely bacteriophage T4, as a model in vivo system for studying dNTP pool imbalances and mutagenesis. While these investigations focus on two enzymes in T4 deoxyribonucleotide metabolism, namely T4 ribonucleotide reductase and dCMP deaminase, the effects of other Cited by: 1.

Deoxyribonucleotides as determinants of DNA replication fidelity in bacteriophage T4. namely bacteriophage T4, as a model in vivo system for studying dNTP pool imbalances and mutagenesis. While these investigations focus on two enzymes in T4 deoxyribonucleotide metabolism, namely T4 ribonucleotide reductase and dCMP deaminase, the.

DEOXYRIBONUCLEOTIDES AS DETERMINANTS OF DNA REPLICATION FIDELITY IN BACTERIOPHAGE T4 I. INTRODUCTION DNA replication results in as few as 1 x to 1 x errors per base pair per round of replication (Drake, ).

In the absence of mutagenic agents during replication, this fidelity is the product. Deoxyribonucleotides. The single-stranded DNA from the structure of the exonuclease domain of T4 bacteriophage (PDB: 1NOY) Determinants of DNA Replication Fidelity. There is a balance between nucleotide selectivity, proofreading efficiency, mismatch repair, nucleotide pools, and the effect of the nature of the DNA template (sequence Cited by: The bacteriophage T4 DNA replication system is a relatively simple system of ten T4 encoded proteins that together catalyze rapid and highly accurate copying of the two strands of a replication.

Since T4 phage-encoded ribonucleotide reductase is insensitive to feedback inhibition, we established conditions under which phage DNA replication is dependent upon ribonucleotide reductase of the host, Escherichia coli. We examined BrdUrd mutagenesis of rII mutants known to revert to wild type either by AT-to-GC or GC-to-AT transition pathways.

The gp59 mediator promotes T4 primosome assembly on single-stranded DNA covered with the gp32 and UvsX proteins. At late times of infection of an E. coli bacterium by T4 bacteriophage, the replication of the viral DNA is initiated by a process that requires genetic recombination (Luder and Mosig, ; Mosig, ).The T4 strand-exchange protein, UvsX, is essential for this synthesis, and.

Construction and characterization of a bacteriophage T4 DNA polymerase deficient in 3'-->5' exonuclease activity. Proc Natl Acad Sci U S A.

Apr 1; 90 (7)– [PMC free article] [Google Scholar] Gauss P, Park K, Spencer TE, Hacker KJ. DNA helicase requirements for DNA replication during bacteriophage T4 infection. J Bacteriol. To understand the molecular basis of mutation stimulated by deoxyribonucleotide pool imbalance, we studied a temperature-sensitive T4 phage gene 42 mutant (LB3), which specifies a thermolabile deoxycytidylate hydroxymethylase.

Analysis of rII mutations, revertible to wild type along either GC-to-AT or AT-to-GC transition pathways, showed 8- to fold stimulation of GC-to-AT mutations at a.

Our laboratory has reported data suggesting a role for T4 phage gene 32 single-stranded DNA-binding protein in organizing a complex of deoxyribonucleotide-synthesizing enzymes at the replication fork. [Show full abstract] bacteriophage T4 revealed essential features of the molecular nature of genes and genomes, mechanism and fidelity of DNA replication, genetic recombination, DNA repair.

Construction and characterization of a bacteriophage T4 DNA polymerase deficient in 3'-->5' exonuclease activity. Proc Natl Acad Sci U S A. Apr 1; 90 (7)– [PMC free article] Gauss P, Park K, Spencer TE, Hacker KJ.

DNA helicase requirements for DNA replication during bacteriophage T4 infection. J Bacteriol. Another interesting point is that another mutant DNA polymerase, the T4 LI-DNA polymerase, has increased replication fidelity.

Thus, conservative amino acid substitutions for L in the Motif A sequence of the T4 DNA polymerase, LM and LI, have profound effects on DNA replication fidelity, which provides a glimpse of the exquisite. The rate of DNA replication in a living cell was first measured as the rate of phage T4 DNA elongation in phage-infected E.

coli. During the period of exponential DNA increase at 37 °C, the rate was nucleotides per second. The mutation rate per base pair per replication during phage T4 DNA. Three groups of T4 DNA polymerase mutants were prepared and characterized.

In the first group, Ala and Asn were substituted for four acidic residues in the exonuclease domain that were chosen on the basis of their sequence alignment with the Klenow fragment from Escherichia coli DNA polymerase I.

Two divalent metal ions required for catalyzing the 3‘−5‘ exonuclease reaction are ligated. Gillin FD, Nossal NG. T4 DNA polymerase has a lower apparent Km for deoxynucleoside triphosphates complementary rather than noncomplementary to the template.

Biochem Biophys Res Commun. May 19; 64 (2)– Liu CC, Burke RL, Hibner U, Barry J, Alberts B. Probing DNA replication mechanisms with the T4 bacteriophage in vitro system.

Kinetic characterization of the polymerase and exonuclease activities of the gene 43 protein of bacteriophage T4. Biochemistry31 (45), DOI: /bia Mark C.

Young, Michael K. Reddy, Peter H. Von Hippel. Structure and function of the bacteriophage T4 DNA polymerase holoenzyme. Changes in other genes involved in replication, including single-stranded DNA-binding proteins and helicase and clamp proteins, can also produce a mutator phenotype in T4, albeit typically more modestly than low-fidelity polymerases.

T4 antimutators showing fold increase in replication fidelity have also been described and often map to the. Motif A of bacteriophage T4 DNA polymerase: role in primer extension and DNA replication fidelity. Isolation of new antimutator and mutator DNA polymerases.

J Biol Chem. ; – Reha-Krantz LJ, Nonay RL, Stocki S. Bacteriophage T4 DNA polymerase mutations that confer sensitivity to the PPi analog phosphonoacetic acid.

J Virol. DNA Repair 4, – Crossref Medline, Google Scholar; Creighton S., Goodman M. () Gel kinetic analysis of DNA polymerase fidelity in the presence of proofreading using bacteriophage T4 DNA polymerase. Biol. Chem. – Crossref Medline, Google Scholar.

Hibner, U. and Alberts, B. M. () Fidelity of DNA replication catalyzed in vitro on a natural DNA template by the T4 bacteriophage multi-enzyme complex. Nature. L.J. Reha-Krantz, in Brenner's Encyclopedia of Genetics (Second Edition), Discontinuous Replication Generates Okazaki Fragments.

Okazaki fragments in bacteria and in bacteriophage T4 are – nucleotides long, but are only about – nucleotides in eukaryotes. Because DNA polymerases cannot initiate DNA synthesis, each Okazaki fragment is primed with a short RNA.In bacteriophage T4 the protein product of gene 43 (gp43) is a multifunctional DNA polymerase that is essential for replication of the phage genome.