Document Detail

Locking the DNA gate of DNA gyrase: investigating the effects on DNA cleavage and ATP hydrolysis.
MedLine Citation:
PMID:  10571989     Owner:  NLM     Status:  MEDLINE    
Supercoiling by DNA gyrase involves the passage of one segment of double-stranded DNA through another. This requires a DNA duplex to be cleaved and the broken ends separated by at least 20 A. This is accomplished by the opening of a dimer interface, termed the DNA gate, which is covalently attached to the broken ends of the DNA. After strand passage, the DNA gate closes allowing the reunion of the broken ends. We have cross-linked the DNA gate of gyrase using cysteine cross-linking to block gate opening. We show that this locked gate mutant can bind quinolone drugs and perform DNA cleavage. However, locking the DNA gate prevents strand passage and the ability of DNA to stimulate ATP hydrolysis. We discuss the mechanistic implications of these results.
N L Williams; A Maxwell
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemistry     Volume:  38     ISSN:  0006-2960     ISO Abbreviation:  Biochemistry     Publication Date:  1999 Oct 
Date Detail:
Created Date:  1999-12-17     Completed Date:  1999-12-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370623     Medline TA:  Biochemistry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  14157-64     Citation Subset:  IM    
Department of Biochemistry, University of Leicester, UK.
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MeSH Terms
Adenosine Triphosphate / metabolism*
Adenylyl Imidodiphosphate / pharmacology
Calcium / pharmacology
Cross-Linking Reagents / metabolism,  pharmacology
Cysteine / metabolism
DNA Topoisomerases, Type II / antagonists & inhibitors*,  chemistry,  genetics,  metabolism*
DNA, Superhelical / chemistry,  metabolism*
Disulfides / metabolism
Mutagenesis, Site-Directed
Nucleic Acid Conformation
Protein Conformation
Protein Structure, Tertiary
Quinolones / metabolism,  pharmacology
Reg. No./Substance:
0/Cross-Linking Reagents; 0/DNA, Superhelical; 0/Disulfides; 0/Quinolones; 25612-73-1/Adenylyl Imidodiphosphate; 52-90-4/Cysteine; 56-65-5/Adenosine Triphosphate; 7440-70-2/Calcium; EC Topoisomerases, Type II

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