Functional Characterization of the Genes Coding for the Terminal Protein and DNA Polymerase from Bacteriophage GA-1. Evidence for a Sliding-back Mechanism During Protein-primed GA-1 DNA Replication

1996 ◽  
Vol 264 (3) ◽  
pp. 453-464 ◽  
Author(s):  
Belén Illana ◽  
Luis Blanco ◽  
Margarita Salas
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Functional Characterization ◽  
Terminal Protein

scholarly journals Expression, purification and characterization of the Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) DNA polymerase and interaction with the SpliNPV non-hr origin of DNA replication

2001 ◽  
Vol 82 (7) ◽  
pp. 1767-1776 ◽  
Author(s):  
Jianhe Huang ◽  
David B. Levin
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Spodoptera Littoralis ◽  
Expression System ◽  
Functional Characterization ◽  
Eukaryotic Expression ◽  
Replication Assay ◽  
E Coli ◽  
Origin Of Dna Replication

The DNA polymerase from Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) was expressed in, and purified from, prokaryotic and eukaryotic expression systems. While less protein was obtained from the E. coli expression system, SpliNPV DNAPOL purified from E. coli displayed similar biochemical characteristics to DNAPOL expressed in, and subsequently purified from, insect cells (Sf9) using a baculovirus expression system. Biochemical analyses suggested that the DNA polymerase and the 3′–5′ exonuclease activities are intrinsic to the protein. Deletion of the first 80 amino acid residues from the N terminus of the DNAPOL affected neither the DNA polymerase nor the exonuclease activities of the enzyme. Replication products from single-stranded M13 DNA demonstrated that the DNA synthesis activity of SpliNPV DNAPOL is highly processive. Transient expression assays with a set of deletion clones containing the putative SpliNPV non-hr origin of DNA replication permitted functional characterization of sequence elements within the origin fragment. Purified SpliNPV DNAPOL stimulated origin-dependent DNA replication in a cell-free replication assay.

scholarly journals Biochemical and functional characterization of Plasmodium falciparum DNA polymerase δ

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
Jitlada Vasuvat ◽  
Atcha Montree ◽  
Sangduen Moonsom ◽  
Ubolsree Leartsakulpanich ◽  
Songsak Petmitr ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Dna Polymerase ◽  
Functional Characterization ◽  
Dna Polymerase Δ

scholarly journals Functional Requirement of Noncoding Y RNAs for Human Chromosomal DNA Replication

2006 ◽  
Vol 26 (18) ◽  
pp. 6993-7004 ◽  
Author(s):  
Christo P. Christov ◽  
Timothy J. Gardiner ◽  
Dávid Szüts ◽  
Torsten Krude
Keyword(s):  
Dna Replication ◽  
Functional Characterization ◽  
Noncoding Rnas ◽  
Biological Processes ◽  
Replication Forks ◽  
Chromosomal Dna ◽  
Free System ◽  
Specific Degradation

ABSTRACT Noncoding RNAs are recognized increasingly as important regulators of fundamental biological processes, such as gene expression and development, in eukaryotes. We report here the identification and functional characterization of the small noncoding human Y RNAs (hY RNAs) as novel factors for chromosomal DNA replication in a human cell-free system. In addition to protein fractions, hY RNAs are essential for the establishment of active chromosomal DNA replication forks in template nuclei isolated from late-G1-phase human cells. Specific degradation of hY RNAs leads to the inhibition of semiconservative DNA replication in late-G1-phase template nuclei. This inhibition is negated by resupplementation of hY RNAs. All four hY RNAs (hY1, hY3, hY4, and hY5) can functionally substitute for each other in this system. Mutagenesis of hY1 RNA showed that the binding site for Ro60 protein, which is required for Ro RNP assembly, is not essential for DNA replication. Degradation of hY1 RNA in asynchronously proliferating HeLa cells by RNA interference reduced the percentages of cells incorporating bromodeoxyuridine in vivo. These experiments implicate a functional role for hY RNAs in human chromosomal DNA replication.

Immunological characterization of the role of adenovirus terminal protein in viral DNA replication

Virology ◽  
1983 ◽  
Vol 131 (2) ◽  
pp. 287-295 ◽  
Author(s):  
A.W.M. Rijnders ◽  
B.G.M. Van Bergen ◽  
P.C. Van Der Vliet ◽  
J.S. Sussenbach
Keyword(s):  
Dna Replication ◽  
Viral Dna ◽  
Terminal Protein ◽  
Viral Dna Replication ◽  
Immunological Characterization
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