Properties of the human Cdc45/Mcm2-7/GINS helicase complex and its action with DNA polymerase ε in rolling circle DNA synthesis Journal Article


Authors: Kang, Y. H.; Galal, W. C.; Farina, A.; Tappin, I.; Hurwitz, J.
Article Title: Properties of the human Cdc45/Mcm2-7/GINS helicase complex and its action with DNA polymerase ε in rolling circle DNA synthesis
Abstract: In eukaryotes, although the Mcm2-7 complex is a key component of the replicative DNA helicase, its association with Cdc45 and GINS (the CMG complex) is required for the activation of the DNA helicase. Here, we show that the CMG complex is localized to chromatin in human cells and describe the biochemical properties of the human CMG complex purified from baculovirus-infected Sf9 cells. The isolated complex binds to ssDNA regions in the presence of magnesium and ATP (or a nonhydrolyzable ATP analog), contains maximal DNAhelicase in the presence of forked DNA structures, and translocates along the leading strand (3′ to 5′ direction). The complex hydrolyses ATP in the absence of DNA; unwinds duplex regions up to 500 bp; and either replication protein A or Escherichia coli single stranded binding protein increases the efficiency of displacement of long duplex regions. Using a 200-nt primed circular DNA substrate, the combined action of human DNA polymerase ε and the human CMG complex leads to the formation of products >10 kb in length. These findings suggest that the coordinated action of these replication complexes supports leading strand synthesis.
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 109
Issue: 16
ISSN: 0027-8424
Publisher: National Academy of Sciences  
Date Published: 2012-04-17
Start Page: 6042
End Page: 6047
Language: English
DOI: 10.1073/pnas.1203734109
PROVIDER: scopus
PUBMED: 22474384
PMCID: PMC3341058
DOI/URL:
Notes: --- - "Export Date: 1 May 2012" - "CODEN: PNASA" - "Source: Scopus"
Altmetric
Citation Impact
MSK Authors
  1. Andrea Farina
    9 Farina
  2. Young-Hoon Kang
    7 Kang
  3. Jerard Hurwitz
    191 Hurwitz
  4. Inger Tappin
    15 Tappin