A mutational analysis of the Bacillus subtilis competence helicase ComFA
Genetic competence is a developmental process in bacteria that allows natural transformation. Competent Gram positive bacteria such as Bacillus subtilis carry a cytosolic helicase which is required for efficient transformation. In this work ComFA is confirmed as a DEAD-box helicase. I also describ...
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| Language: | en_US |
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Harvard University
2014
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| Online Access: | http://dissertations.umi.com/gsas.harvard:11560 http://nrs.harvard.edu/urn-3:HUL.InstRepos:12274569 |
| Summary: | Genetic competence is a developmental process in bacteria that allows natural transformation. Competent Gram positive bacteria such as Bacillus subtilis carry a cytosolic helicase which is required for efficient transformation. In this work ComFA is confirmed as a DEAD-box helicase. I also describe a new accessory motif in ComFA that contributes to transformation independently of the helicase activity in ComFA. The newly discovered metal-binding motif consists of four cysteines which are required for transformation and zinc binding. While the zinc finger is required for full function, it is not required for DNA binding. As DEAD-box family helicases are generally non-processive, it appears that at least part of the rapid DNA uptake process is mediated by a non-processive helicase. Active uptake using the ComFA helicase motor may be required to maintain the integrity of the incoming DNA to allow subsequent recombination. |
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