A conserved MCM single-stranded DNA binding element is essential for replication initiation
Name
Froelich-2014-A conserved MCM sing.pdf
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4.12 MB
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Author(s)
Froelich, Clifford A.
Kang, Sukhyun
Epling, Leslie B.
Enemark, Eric J.
Bell, Stephen P
Date Issued
April 2014
Journal
eLife
Publisher
eLife Sciences Publications, Ltd.
Citation
Froelich, Clifford A, Sukhyun Kang, Leslie B Epling, Stephen P Bell, and Eric J Enemark. “A Conserved MCM Single-Stranded DNA Binding Element Is Essential for Replication Initiation.” eLife 3 (April 1, 2014). pp 1-21.
Version
Final published version
Abstract
The ring-shaped MCM helicase is essential to all phases of DNA replication. The complex loads at replication origins as an inactive double-hexamer encircling duplex DNA. Helicase activation converts this species to two active single hexamers that encircle single-stranded DNA (ssDNA). The molecular details of MCM DNA interactions during these events are unknown. We determined the crystal structure of the Pyrococcus furiosus MCM N-terminal domain hexamer bound to ssDNA and define a conserved MCM-ssDNA binding motif (MSSB). Intriguingly, ssDNA binds the MCM ring interior perpendicular to the central channel with defined polarity. In eukaryotes, the MSSB is conserved in several Mcm2-7 subunits, and MSSB mutant combinations in S. cerevisiae Mcm2-7 are not viable. Mutant Mcm2-7 complexes assemble and are recruited to replication origins, but are defective in helicase loading and activation. Our findings identify an important MCM-ssDNA interaction and suggest it functions during helicase activation to select the strand for translocation.
MIT Department
Massachusetts Institute of Technology. Department of Biology
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DOI of Published Version
https://doi.org/10.7554/eLife.01993
