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DNA aggregation by an archaeal DNA binding protein Sac10b and its novel DNA nicking activity

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Title DNA aggregation by an archaeal DNA binding protein Sac10b and its novel DNA nicking activity
 
Creator Koti, Jaya S
Kanugula, Sreenivas
Suryanarayana, Tangirala
 
Subject Sulfolobus acidocaldarius
Sac10b
DNA aggregation
DNA nicking
 
Description 166-173
The solution structure of an archaeal DNA binding protein Sac10b (DBNP-B) by cross-linking with formaldehyde and its
interaction with DNA were studied. Results indicated that Sac10b existed as oligomeric structure in solution and the
oligomerization was greatly stimulated by Mg²⁺ ions and elevated temperatures. In light of our earlier observation that
Sac10b interacts with DNA forming different types of complexes with DNA at different protein concentrations, its DNAbinding
properties were also studied. Results demonstrated that the protein formed rapidly sedimentable
co-aggregation complexes with both native and denatured DNA in a protein concentration-dependent manner. These protein
DNA complexes were fluid-like crystalline material at a protein DNA ratio (3-6:1 w/w). Gel mobility shift assays carried
out to study the interaction of the protein with plasmid DNA indicated possible DNA nicking by the protein. The DNA
nicking activity of Sac10b was optimal in pH range of 7-8.5 and was dependent on Mg²⁺ ions. It was maximal at protein to
DNA ratio of (8:1, w/w) and very little activity was observed above and below this ratio. Nicking of DNA at this ratio
indicated structure-specific DNA nicking by the protein. The protein might have important multi-functional role in the DNA
metabolism in this organism.
 
Date 2008-08-29T11:10:15Z
2008-08-29T11:10:15Z
2008-06
 
Type Article
 
Identifier 0301-1208
http://hdl.handle.net/123456789/1928
 
Language en_US
 
Publisher CSIR
 
Source IJBB Vol.45(3) [June 2008]