Journal article
Polarity of human replication protein A binding to DNA
Nucleic acids research, Vol.29(2), pp.373-379
01/15/2001
DOI: 10.1093/nar/29.2.373
PMCID: PMC29666
PMID: 11139606
Abstract
Replication protein A (RPA), the nuclear single-stranded DNA
binding protein is involved in DNA replication, nucleotide excision
repair (NER) and homologous recombination. It is a stable heterotrimer
consisting of subunits with molecular masses of 70, 32 and 14 kDa
(p70, p32 and p14, respectively). Gapped DNA structures are common
intermediates during DNA replication and NER. To analyze the interaction
of RPA and its subunits with gapped DNA we designed structures containing
9 and 30 nucleotide gaps with a photoreactive arylazido
group at the 3′-end of the upstream
oligonucleotide or at the 5′-end of the
downstream oligonucleotide. UV crosslinking and subsequent analysis
showed that the p70 subunit mainly interacts with the 5′-end
of DNA irrespective of DNA structure, while the subunit orientation
towards the 3′-end of DNA in the gap
structures strongly depends on the gap size. The results are compared
with the data obtained previously with the primer–template systems
containing 5′- or 3′-protruding
DNA strands. Our results suggest a model of polar RPA binding to the
gapped DNA.
Details
- Title: Subtitle
- Polarity of human replication protein A binding to DNA
- Creators
- Dmitry M Kolpashchikov - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, RussiaSvetlana N Khodyreva - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, RussiaDenis Yu Khlimankov - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, RussiaMarc S Wold - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, RussiaAlain Favre - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, RussiaOlga I Lavrik - Novosibirsk Institute of Bioorganic Chemistry, Siberian Division of Russian Academy of Sciences, Prospect Lavrentiev 8, 630090, Novosibirsk, Russia
- Resource Type
- Journal article
- Publication Details
- Nucleic acids research, Vol.29(2), pp.373-379
- Publisher
- Oxford University Press; Oxford, UK
- DOI
- 10.1093/nar/29.2.373
- PMID
- 11139606
- PMCID
- PMC29666
- ISSN
- 0305-1048
- eISSN
- 1362-4962
- Language
- English
- Date published
- 01/15/2001
- Academic Unit
- Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024549102771
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