Mechanistic analysis of local ori melting and helicase assembly by the papillomavirus e1 protein

Schuck, S., Stenlund, A. (September 2011) Mechanistic analysis of local ori melting and helicase assembly by the papillomavirus e1 protein. Mol Cell, 43 (5). pp. 776-787. ISSN 1097-4164 (Electronic) 1097-2765 (Linking)

Abstract

Preparation of DNA templates for replication requires opening of the duplex to expose single-stranded (ss) DNA. The locally melted DNA is required for replicative DNA helicases to initiate unwinding. How local melting is generated in eukaryotic replicons is unknown, but initiator proteins from a handful of eukaryotic viruses can perform this function. Here we dissect the local melting process carried out by the papillomavirus E1 protein. We characterize the melting process kinetically and identify mutations in the E1 helicase and in the ori that arrest the local melting process. We show that a subset of these mutants have specific defects for melting of the center of the ori containing the binding sites for E1 and demonstrate that these mutants fail to untwist the ori DNA. This understanding of how E1 generates local melting suggests possible mechanisms for local melting in other replicons.

Item Type: Paper
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 > enzymes > helicase
organism description > virus
CSHL Authors:
Communities: CSHL Cancer Center Shared Resources > DNA Sequencing Service
CSHL labs > Stenlund lab
CSHL Cancer Center Program > Gene Regulation and Cell Proliferation
Depositing User: Brian Soldo
Date: 2 September 2011
Date Deposited: 14 Mar 2012 15:42
Last Modified: 13 Oct 2015 16:20
PMCID: PMC3209524
Related URLs:
URI: https://repository.cshl.edu/id/eprint/25173

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