Characterization of the function of yeast Rsc8 SWIRM domain
| Autor: | Chin-Yu Chang, 張瑾愉 |
|---|---|
| Rok vydání: | 2009 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 97 Yeast RSC complex contains seventeen subunits with the capacity to remodel the structure of chromatin. Rsc8p, a 64 kDa protein, contains a conservative SWIRM domain named after the proteins Swi3, Rsc8 and Moira in which it was first recognized. To investigate the role of the SWIRM domain on chromatin remodeling and its consequent cellular functions, five rsc8 temperature sensitive mutants (ts mutants) with mutations on the SWIRM domain were generated by PCR mutagenesis. In further characterization, four out of five SWIRM domain ts mutants were found to be sensitive to phleomycin and benomyl treatment. Further, yeast two-hybrid screening approach was used to isolate proteins that interact with the SWIRM domain of Rsc8 protein. Two proteins, Rsc7, a component of RSC complex known to interact with Rsc8, and a novel yeast RING protein, Cwc24, involved in mRNA splicing were isolated. In all but SWIRM77 mutants demonstrated decreased interactions between mutant SWIRM domain and Rsc7, and the SWIRM77 showed reduced interactions between Cwc24. The in vivo interaction between Rsc8 and Cwc24 was not observed by Co-immunoprecipitation. In combination of protein interaction and mutant phenotypes, mutation sites, molecular modeling, SWIRM domain can be divided into two functional regions each interacted with Rsc7 and Cwc24. The interaction between Rsc7 may be related to the structure integrity of RSC complex, and the interaction between Cwc24 may suggest the functional relationship between chromatin remodeling and RNA processing. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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