Document Detail


Sin resolvase catalytic activity and oligomerization state are tightly coupled.
MedLine Citation:
PMID:  20868695     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Serine recombinases promote specific DNA rearrangements by a cut-and-paste mechanism that involves cleavage of all four DNA strands at two sites recognized by the enzyme. Dissecting the order and timing of these cleavage events and the steps leading up to them is difficult because the cleavage reaction is readily reversible. Here, we describe assays using activated Sin mutants and a DNA substrate with a 3'-bridging phosphorothiolate modification that renders Sin-mediated DNA cleavage irreversible. We find that activating Sin mutations promote DNA cleavage rather than simply stabilize the cleavage product. Cleavage events at the scissile phosphates on complementary strands of the duplex are tightly coupled, and the overall DNA cleavage rate is strongly dependent on Sin concentration. When combined with analytical ultracentrifugation data, these results suggest that Sin catalytic activity and oligomerization state are tightly linked, and that activating mutations promote formation of a cleavage-competent oligomeric state that is normally formed only transiently within the full synaptic complex.
Authors:
Kent W Mouw; Andrew M Steiner; Rodolfo Ghirlando; Nan-Sheng Li; Sally-J Rowland; Martin R Boocock; W Marshall Stark; Joseph A Piccirilli; Phoebe A Rice
Related Documents :
1586945 - Dna cleavage in trans by the active site tyrosine during flp recombination: switching p...
20537435 - Chiral preference of l-tryptophan derived metal-based antitumor agent of late 3d-metal ...
8755245 - The biochemistry of polyadenylation.
9311555 - Interaction of the antitumour antibiotic streptonigrin with dna and oligonucleotides.
17588915 - Comment on "tequila, a neurotrypsin ortholog, regulates long-term memory formation in d...
6280995 - Nucleotide sequence of the simian virus 40 hindii + iii restriction fragment i (fourth ...
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-09-22
Journal Detail:
Title:  Journal of molecular biology     Volume:  404     ISSN:  1089-8638     ISO Abbreviation:  J. Mol. Biol.     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-11-03     Completed Date:  2010-12-06     Revised Date:  2014-09-17    
Medline Journal Info:
Nlm Unique ID:  2985088R     Medline TA:  J Mol Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  16-33     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier Ltd. All rights reserved.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Bacterial Proteins / genetics,  metabolism*
DNA / chemical synthesis,  metabolism*
DNA Nucleotidyltransferases / genetics,  metabolism*
Models, Molecular
Mutant Proteins / genetics,  metabolism
Oligonucleotides / chemical synthesis,  metabolism
Protein Multimerization*
Protein Structure, Quaternary
Recombination, Genetic*
Ultracentrifugation
Grant Support
ID/Acronym/Agency:
GM086826/GM/NIGMS NIH HHS; R01 GM086826/GM/NIGMS NIH HHS; R01 GM086826-02/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/Mutant Proteins; 0/Oligonucleotides; 9007-49-2/DNA; EC 2.7.7.-/DNA Nucleotidyltransferases; EC 2.7.7.-/sin protein, Staphylococcus aureus
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  A simple definition of structural regions in proteins and its use to analyse interface evolution.
Next Document:  Reversibility and efficiency in coding protein information.