Mutational analysis of the central channel in the Simian virus 40 large T antigen helicase
I report in this dissertation that residues lining the small tier of the central channel have an important role in DNA binding, especially in single-stranded DNA binding. Defects in helicase activity for the small tier channel mutants can be accounted by their corresponding defects in DNA binding. O...
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| Format: | Dissertation |
| Language: | English |
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ProQuest Dissertations & Theses
01-01-2006
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| Online Access: | Get full text |
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| Summary: | I report in this dissertation that residues lining the small tier of the central channel have an important role in DNA binding, especially in single-stranded DNA binding. Defects in helicase activity for the small tier channel mutants can be accounted by their corresponding defects in DNA binding. One of the large tier central channel mutants (R456A) showed a defect in unwinding of origin DNA, but this could not be attributed to a problem in DNA binding. Combined with the results of others, it appears that the large tier is primarily involved in unwinding origin DNA. These findings are incorporated into a model of T antigen helicase activity where single-stranded DNA is looped out of one hexamer through hydrophilic side-channels in the large tier of the hexamer. The looped-out single-stranded DNA runs back into the origin binding domain of the adjacent hexamer. In this model, there is one strand of DNA within each small tier during DNA unwinding from the origin. (Abstract shortened by UMI.) |
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| ISBN: | 0542719339 9780542719332 |