Source:http://linkedlifedata.com/resource/pubmed/id/12637557
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
21
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| pubmed:dateCreated |
2003-5-19
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| pubmed:abstractText |
Structural differences between class A and B DNA polymerases suggest that the motif B region, a wall of the catalytic pocket, may have evolved differentially in the two polymerase families. This study examines the function of the motif B residues in Saccharomyces cerevisiae DNA polymerase alpha (pol alpha). Effects of the mutations were determined by biochemical analysis and genetic complementation of a yeast strain carrying a temperature-sensitive pol alpha mutant. Many conserved residues were viable with a variety of substitutions. Among them, mutations at Asn-948 or Tyr-951 conferred up to 8-fold higher colony formation frequency in a URA3 forward mutation assay, and 79-fold higher trp1 reversion frequency was observed for Y951P in yeast. Purified Y951P was as accurate as wild type in DNA synthesis but approximately 6-fold less processive and 22-fold less active in vitro. Therefore, Y951P may increase the frequency of mutant colony formation because of its low level of DNA polymerase activity in yeast. Mutations at Lys-944 or Gly-952 were not viable, which is consistent with the observation that mutants with substitutions at Gly-952 have strongly reduced catalytic activity in vitro. Gly-952 may provide a space for the nascent base pair and thus may play an essential function in S. cerevisiae DNA pol alpha. These results suggest that class B DNA polymerases have a unique structure in the catalytic pocket, which is distinct from the corresponding region in class A DNA polymerases.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/2'-deoxyguanosine 5'-phosphate,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Fungal,
http://linkedlifedata.com/resource/pubmed/chemical/DNA Polymerase I,
http://linkedlifedata.com/resource/pubmed/chemical/Deoxyguanine Nucleotides,
http://linkedlifedata.com/resource/pubmed/chemical/Glycine,
http://linkedlifedata.com/resource/pubmed/chemical/Lysine,
http://linkedlifedata.com/resource/pubmed/chemical/Taq Polymerase
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| pubmed:status |
MEDLINE
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| pubmed:month |
May
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| pubmed:issn |
0021-9258
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
23
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| pubmed:volume |
278
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
19071-8
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:12637557-Amino Acid Sequence,
pubmed-meshheading:12637557-Base Pairing,
pubmed-meshheading:12637557-Binding Sites,
pubmed-meshheading:12637557-Catalysis,
pubmed-meshheading:12637557-Conserved Sequence,
pubmed-meshheading:12637557-DNA, Fungal,
pubmed-meshheading:12637557-DNA Polymerase I,
pubmed-meshheading:12637557-Deoxyguanine Nucleotides,
pubmed-meshheading:12637557-Gene Deletion,
pubmed-meshheading:12637557-Gene Library,
pubmed-meshheading:12637557-Glycine,
pubmed-meshheading:12637557-Kinetics,
pubmed-meshheading:12637557-Lysine,
pubmed-meshheading:12637557-Molecular Sequence Data,
pubmed-meshheading:12637557-Mutagenesis, Insertional,
pubmed-meshheading:12637557-Mutagenesis, Site-Directed,
pubmed-meshheading:12637557-Mutation,
pubmed-meshheading:12637557-Saccharomyces cerevisiae,
pubmed-meshheading:12637557-Sequence Alignment,
pubmed-meshheading:12637557-Structure-Activity Relationship,
pubmed-meshheading:12637557-Taq Polymerase
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| pubmed:year |
2003
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| pubmed:articleTitle |
Distinct function of conserved amino acids in the fingers of Saccharomyces cerevisiae DNA polymerase alpha.
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| pubmed:affiliation |
Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University Graduate School of Medicine, Nagoya 466-8550, Japan.
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| pubmed:publicationType |
Journal Article,
Comparative Study
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