Reconstitution of functional human single-stranded DNA-binding protein from individual subunits expressed by recombinant baculoviruses Journal Article

   


Authors: Stigger, E.; Dean, F. B.; Hurwitz, J.; Lee, S. H.
Article Title: Reconstitution of functional human single-stranded DNA-binding protein from individual subunits expressed by recombinant baculoviruses
Abstract: Human single-stranded DNA-binding protein (HSSB), also known as replication protein A, is composed of a 70-kDa single-stranded DNA-binding subunit (p70) and 34-kDa and 11-kDa (p34 and p11, respectively) subunits of unknown functions. We have examined interactions among the HSSB subunits in vivo by coinfecting insect cells with different combinations of recombinant baculoviruses encoding p70, p34, or p11. In vivo, coexpressed p34 and p11 subunits formed stable complexes, whereas neither p34 nor p11 formed stable complexes with p70. In cells coinfected with viruses expressing all three subunits, the stable heterotrimer formed, which, when purified, replaced HSSB isolated from HeLa cells in various assays, including simian virus 40 DNA replication in vitro. These data suggest that, in the assembly of functionally active HSSB, formation of the p34-p11 complex precedes p70 addition to the complex.
Keywords: dna binding protein; nonhuman; dna polymerase; dna replication; complex formation; hela cell; protein processing; dna viruses; simian virus 40; virus replication; single stranded dna; cell interaction; virus expression; virus isolation; insect; enzyme reconstitution; baculovirus; protein quaternary structure; simiae; unidentified baculovirus; insecta; simian virus; priority journal; article
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 91
Issue: 2
ISSN: 0027-8424
Publisher: National Academy of Sciences  
Date Published: 1994-01-18
Start Page: 579
End Page: 583
Language: English
DOI: 10.1073/pnas.91.2.579
PROVIDER: scopus
PMCID: PMC42992
PUBMED: 8290566
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-0028157927&doi=10.1073%2fpnas.91.2.579&partnerID=40&md5=09fbb9d496fc2abf57a32aa8727ae4a5
Notes: Export Date: 14 January 2019 -- Article -- CODEN: PNASA C2 -- Source: Scopus
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MSK Authors
  1. Jerard Hurwitz
    206 Hurwitz
  2. Frank B. Dean
    23 Dean
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