Kinetics of DNA Polymerase I (Klenow Fragment Exo-) Activity on Damaged DNA Templates:  Effect of Proximal and Distal Template Damage on DNA Synthesis†

Biochemistry ◽  
1997 ◽  
Vol 36 (49) ◽  
pp. 15336-15342 ◽  
Author(s):  
Holly Miller ◽  
Arthur P. Grollman
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Klenow Fragment ◽  
Dna Polymerase I ◽  
Dna Templates ◽  
Polymerase I ◽  
Kinetics Of ◽  
Damaged Dna

scholarly journals DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (12) ◽  
pp. 2811-2817 ◽  
Author(s):  
M A Resnick ◽  
A Sugino ◽  
J Nitiss ◽  
T Chow
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
High Efficiency ◽  
Intragenic Recombination ◽  
Dna Polymerase I ◽  
Crude Extracts ◽  
Total Dna ◽  
Polymerase I ◽  
Kinetics Of

We utilized strains of Saccharomyces cerevisiae that exhibit high efficiency of synchrony of meiosis to examine several aspects of meiosis including sporulation, recombination, DNA synthesis, DNA polymerase I and II, and Mg2+-dependent alkaline DNases. The kinetics of commitment to intragenic recombination and sporulation are similar. The synthesis of DNA, as measured directly with diphenylamine, appears to precede the commitment to recombination. Both DNA polymerase I and II activities and total DNA-synthesizing activity in crude extracts increase two- to threefold before the beginning of meiotic DNA synthesis. Increases of 10- to 20-fold over mitotic levels are found for Mg2+-dependent alkaline DNase activity in crude extracts before and during the commitment to meiotic intragenic recombination. Of particular interest is the comparable increase in a nuclease under the control of the RAD52 gene; this enzyme has been identified by the use of antibody raised against a similar enzyme from Neurospora crassa. Since the RAD52 gene is essential for meiotic recombination, the nuclease is implicated in the high levels of recombination observed during meiosis. The effects observed in this report are meiosis specific since they are not observed in an alpha alpha strain.

DNA polymerases, deoxyribonucleases, and recombination during meiosis in Saccharomyces cerevisiae

1984 ◽  
Vol 4 (12) ◽  
pp. 2811-2817
Author(s):  
M A Resnick ◽  
A Sugino ◽  
J Nitiss ◽  
T Chow
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
High Efficiency ◽  
Intragenic Recombination ◽  
Dna Polymerase I ◽  
Crude Extracts ◽  
Total Dna ◽  
Polymerase I ◽  
Kinetics Of

We utilized strains of Saccharomyces cerevisiae that exhibit high efficiency of synchrony of meiosis to examine several aspects of meiosis including sporulation, recombination, DNA synthesis, DNA polymerase I and II, and Mg2+-dependent alkaline DNases. The kinetics of commitment to intragenic recombination and sporulation are similar. The synthesis of DNA, as measured directly with diphenylamine, appears to precede the commitment to recombination. Both DNA polymerase I and II activities and total DNA-synthesizing activity in crude extracts increase two- to threefold before the beginning of meiotic DNA synthesis. Increases of 10- to 20-fold over mitotic levels are found for Mg2+-dependent alkaline DNase activity in crude extracts before and during the commitment to meiotic intragenic recombination. Of particular interest is the comparable increase in a nuclease under the control of the RAD52 gene; this enzyme has been identified by the use of antibody raised against a similar enzyme from Neurospora crassa. Since the RAD52 gene is essential for meiotic recombination, the nuclease is implicated in the high levels of recombination observed during meiosis. The effects observed in this report are meiosis specific since they are not observed in an alpha alpha strain.

scholarly journals Crystallization and preliminary X-ray analysis of thePlasmodium falciparumapicoplast DNA polymerase

2015 ◽  
Vol 71 (3) ◽  
pp. 333-337 ◽  
Author(s):  
Morgan E. Milton ◽  
Jun-yong Choe ◽  
Richard B. Honzatko ◽  
Scott W. Nelson
Keyword(s):  
Dna Polymerase ◽  
Structural Information ◽  
Klenow Fragment ◽  
X Ray Diffraction ◽  
Dna Polymerase I ◽  
X Ray ◽  
Cell Parameters ◽  
A Genome ◽  
Reported Data ◽  
Polymerase I

Infection by the parasitePlasmodium falciparumis the leading cause of malaria in humans. The parasite has a unique and essential plastid-like organelle called the apicoplast. The apicoplast contains a genome that undergoes replication and repair through the action of a replicative polymerase (apPOL). apPOL has no direct orthologs in mammalian polymerases and is therefore an attractive antimalarial drug target. No structural information exists for apPOL, and the Klenow fragment ofEscherichia coliDNA polymerase I, which is its closest structural homolog, shares only 28% sequence identity. Here, conditions for the crystallization of and preliminary X-ray diffraction data from crystals ofP. falciparumapPOL are reported. Data complete to 3.5 Å resolution were collected from a single crystal (2 × 2 × 5 µm) using a 5 µm beam. The space groupP6522 (unit-cell parametersa=b= 141.8,c= 149.7 Å, α = β = 90, γ = 120°) was confirmed by molecular replacement. Refinement is in progress.

scholarly journals Crystal structures of DNA polymerase I capture novel intermediates in the DNA synthesis pathway

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Nicholas Chim ◽  
Lynnette N Jackson ◽  
Anh M Trinh ◽  
John C Chaput
Keyword(s):  
High Resolution ◽  
Dna Synthesis ◽  
Crystal Structures ◽  
Dna Polymerase ◽  
Active Site ◽  
Dna Polymerase I ◽  
Dynamic Nature ◽  
Enzyme Active Site ◽  
Polymerase I ◽  
In Cells

High resolution crystal structures of DNA polymerase intermediates are needed to study the mechanism of DNA synthesis in cells. Here we report five crystal structures of DNA polymerase I that capture new conformations for the polymerase translocation and nucleotide pre-insertion steps in the DNA synthesis pathway. We suggest that these new structures, along with previously solved structures, highlight the dynamic nature of the finger subdomain in the enzyme active site.

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