Document Detail

Structural insights into the adaptation of proliferating cell nuclear antigen (PCNA) from Haloferax volcanii to a high-salt environment.
MedLine Citation:
PMID:  19770505     Owner:  NLM     Status:  MEDLINE    
The sliding clamp proliferating cell nuclear antigen (PCNA) plays vital roles in many aspects of DNA replication and repair in eukaryotic cells and in archaea. Realising the full potential of archaea as a model for PCNA function requires a combination of biochemical and genetic approaches. In order to provide a platform for subsequent reverse genetic analysis, PCNA from the halophilic archaeon Haloferax volcanii was subjected to crystallographic analysis. The gene was cloned and expressed in Escherichia coli and the protein was purified by affinity chromatography and crystallized by the vapour-diffusion technique. The structure was determined by molecular replacement and refined at 3.5 A resolution to a final R factor of 23.7% (R(free) = 25%). PCNA from H. volcanii was found to be homotrimeric and to resemble other homotrimeric PCNA clamps but with several differences that appear to be associated with adaptation of the protein to the high intracellular salt concentrations found in H. volcanii cells.
Ekaterina Morgunova; Fiona C Gray; Stuart A Macneill; Rudolf Ladenstein
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-09-16
Journal Detail:
Title:  Acta crystallographica. Section D, Biological crystallography     Volume:  65     ISSN:  1399-0047     ISO Abbreviation:  Acta Crystallogr. D Biol. Crystallogr.     Publication Date:  2009 Oct 
Date Detail:
Created Date:  2009-09-22     Completed Date:  2010-01-26     Revised Date:  2013-03-05    
Medline Journal Info:
Nlm Unique ID:  9305878     Medline TA:  Acta Crystallogr D Biol Crystallogr     Country:  England    
Other Details:
Languages:  eng     Pagination:  1081-8     Citation Subset:  IM    
Karolinska Institutet, NOVUM, Centre of Structural Biochemistry, S-14157 Huddinge, Sweden.
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MeSH Terms
Amino Acid Sequence
Cloning, Molecular
Crystallography, X-Ray*
Escherichia coli / genetics
Haloferax volcanii / chemistry*,  metabolism
Models, Molecular
Molecular Sequence Data
Proliferating Cell Nuclear Antigen / chemistry*,  genetics,  metabolism
Salts / metabolism
Sequence Alignment
Reg. No./Substance:
0/Proliferating Cell Nuclear Antigen; 0/Salts

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