ATPase-dependent cooperative binding of ORC and Cdc6 to origin DNA

Speck, C., Chen, Z., Li, H., Stillman, B. (2005) ATPase-dependent cooperative binding of ORC and Cdc6 to origin DNA. Nat Struct Mol Biol, 12 (11). pp. 965-71. ISSN 1545-9993 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/16228006
DOI: 10.1038/nsmb1002

Abstract

Binding of Cdc6 to the origin recognition complex (ORC) is a key step in the assembly of a pre-replication complex (pre-RC) at origins of DNA replication. ORC recognizes specific origin DNA sequences in an ATP-dependent manner. Here we demonstrate cooperative binding of Saccharomyces cerevisiae Cdc6 to ORC on DNA in an ATP-dependent manner, which induces a change in the pattern of origin binding that requires the Orc1 ATPase. The reaction is blocked by specific origin mutations that do not interfere with the interaction between ORC and DNA. Single-particle reconstruction of electron microscopic images shows that the ORC-Cdc6 complex forms a ring-shaped structure with dimensions similar to those of the ring-shaped MCM helicase. The ORC-Cdc6 structure is predicted to contain six AAA+ subunits, analogous to other ATP-dependent protein machines. We suggest that Cdc6 and origin DNA activate a molecular switch in ORC that contributes to pre-RC assembly.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Cell Cycle Proteins metabolism/ultrastructure DNA genetics metabolism DNA Footprinting DNA Primers Electrophoretic Mobility Shift Assay Microscopy Electron Models Molecular Molecular Sequence Data Multiprotein Complexes chemistry ultrastructure Mutation genetics Origin Recognition Complex metabolism ultrastructure Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins metabolism ultrastructure Structure Activity Relationship
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > origin recognition complex
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 05 Jan 2012 14:23
Last Modified: 20 Jun 2017 18:12
PMCID: PMC2952294
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22712

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving