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RPA subunit arrangement near the 3'-end of the primer is modulated by the length of the template strand and cooperative protein interactions.
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MedLine Citation:
PMID:  10518616     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To analyze the interaction of human replication protein A (RPA) and its subunits with the DNA template-primer junction in the DNA replication fork, we designed several template-primer systems differing in the size of the single-stranded template tail (4, 9, 13, 14, 19 and 31 nt). Base substituted photoreactive dNTP analogs-5-[ N -(2-nitro-5-azidobenzoyl)- trans -3-amino-propenyl-1]-2'-deoxyuridine-5'-triphosphate (NAB-4-dUTP) and 5-[ N -[ N -(2-nitro-5-azidobenzoyl)glycyl]- trans -3-aminopropenyl-1]-2'-deoxyuridine-5'-triphosphate (NAB-7-dUTP)-were used as substrates for elongation of radiolabeled primer-template by DNA polymerases in the presence or absence of RPA. Subsequent UV crosslinking showed that the pattern of p32 and p70 RPA subunit labeling, and consequently their interaction with the template-primer junction, is strongly dependent on the template extension length at a particular RPA concentration, as well as on the ratio of RPA to template concentration. Our results suggest a model of changes in the RPA configuration modulating by the length of the template extension in the course of nascent DNA synthesis.
Authors:
O I Lavrik; D M Kolpashchikov; K Weisshart; H P Nasheuer; S N Khodyreva; A Favre
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nucleic acids research     Volume:  27     ISSN:  1362-4962     ISO Abbreviation:  Nucleic Acids Res.     Publication Date:  1999 Nov 
Date Detail:
Created Date:  1999-12-02     Completed Date:  1999-12-02     Revised Date:  2008-11-20    
Medline Journal Info:
Nlm Unique ID:  0411011     Medline TA:  Nucleic Acids Res     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  4235-40     Citation Subset:  IM    
Affiliation:
Novosibirsk Institute of Bioorganic Chemistry Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090 Novosibirsk, Russia. lavrik@niboch.nsc.ru
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MeSH Terms
Descriptor/Qualifier:
Azides / chemistry,  metabolism
Cross-Linking Reagents / chemistry,  metabolism
DNA / biosynthesis
DNA Helicases / chemistry,  metabolism
DNA Polymerase I / metabolism
DNA Polymerase beta / metabolism
DNA Primers / genetics,  metabolism*
DNA Replication / genetics*
DNA, Single-Stranded / chemistry*,  genetics,  metabolism*
DNA-Binding Proteins / chemistry*,  metabolism*
Escherichia coli / enzymology
Humans
Models, Biological
Molecular Weight
Protein Binding
Protein Conformation
Replication Protein A
Templates, Genetic
Ultraviolet Rays
Uridine Triphosphate / analogs & derivatives,  chemistry,  metabolism
Chemical
Reg. No./Substance:
0/5-(N-(2-nitro-5-azidobenzoyl)-3-aminopropen-1-yl)deoxyuridine-5'-triphosphate; 0/Azides; 0/Cross-Linking Reagents; 0/DNA Primers; 0/DNA, Single-Stranded; 0/DNA-Binding Proteins; 0/RPA1 protein, human; 0/Replication Protein A; 63-39-8/Uridine Triphosphate; 9007-49-2/DNA; EC 2.7.7.-/DNA Polymerase I; EC 2.7.7.-/DNA Polymerase beta; EC 3.6.1.-/DNA Helicases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Full Text
Journal Information
Journal ID (nlm-ta): Nucleic Acids Res
ISSN: 0305-1048
ISSN: 1362-4962
Article Information
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Print publication date: Day: 1 Month: 11 Year: 1999
Volume: 27 Issue: 21
First Page: 4235 Last Page: 4240
ID: 148699
PubMed Id: 10518616
Publisher Item Identifier: gkc622

RPA subunit arrangement near the 3'-end of the primer is modulated by the length of the template strand and cooperative protein interactions.
O I Lavrik
D M Kolpashchikov
K Weisshart
H P Nasheuer
S N Khodyreva
A Favre
Novosibirsk Institute of Bioorganic Chemistry Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090 Novosibirsk, Russia. lavrik@niboch.nsc.ru


Article Categories:
  • Research Article


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