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PMID: 12554672 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A model for dsDNA translocation revealed by a structural motif common to RecG and Mfd proteins.

The EMBO journal ·Vol. 22 ·No. 3 ·2003-02-03 ·Pages 724-34

Mahdi AA, Briggs GS, Sharples GJ, Wen Q, Lloyd RG

Abstract

RecG protein differs from other helicases analysed to atomic resolution in that it mediates strand separation via translocation on double-stranded (ds) rather than single-stranded (ss) DNA. We describe a highly conserved helical hairpin motif in RecG and show it to be important for helicase activity. It places two arginines (R609 and R630) in opposing positions within the component helices where they are stabilized by a network of hydrogen bonds involving a glutamate from helicase motif VI. We suggest that disruption of this feature, triggered by ATP hydrolysis, moves an adjacent loop structure in the dsDNA-binding channel and that a swinging arm motion of this loop drives translocation. Substitutions that reverse the charge at R609 or R630 reduce DNA unwinding and ATPase activities, and increase dsDNA binding, but do not affect branched DNA binding. Sequences forming the helical hairpin and loop structures are highly conserved in Mfd protein, a transcription-coupled DNA repair factor that also translocates on dsDNA. The possibility of type I restriction enzymes and chromatin-remodelling factors using similar structures to drive translocation on dsDNA is discussed.

MeSH Terms
Adenosine Triphosphate/metabolism Amino Acid Sequence Arginine/metabolism Bacterial Proteins/chemistry,metabolism DNA/metabolism DNA Helicases/chemistry,metabolism DNA Replication/physiology DNA-Binding Proteins/genetics,metabolism Escherichia coli Proteins/chemistry,genetics,metabolism Hydrogen Bonding Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Structure, Secondary Protein Structure, Tertiary Sequence Alignment Transcription Factors/chemistry,metabolism
Chemicals
Bacterial Proteins DNA-Binding Proteins Escherichia coli Proteins Transcription Factors transcription repair coupling factor protein, Bacteria RecG protein, E coli Adenosine Triphosphate DNA Arginine DNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Mahdi Akeel A
Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK.
Briggs Geoffrey S
Sharples Gary J
Wen Qin
Lloyd Robert G
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdg043
Published
2003-02-03
Pages
724-34
Language
English
Region
England
NLM ID
8208664
PMCID
PMC140728
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/cdg043 PMC Full Text View journal details
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