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Purification and characterization of DNA-dependent RNA polymerase II from Candida utilis

Patturajan, M (1995) Purification and characterization of DNA-dependent RNA polymerase II from Candida utilis. In: Biochemistry & Molecular Biology International, 37 (2). pp. 295-304.

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DNA-dependent RNA polymerase II from Candida utilis has been purified to near homogeneity. The purified enzyme resolved into three subforms, viz. IIO, IIA and IIB. On SDS-PAGE the enzyme showed ten polypeptides with molecular weights in the range of 205 kDa to 14 kDa. By two dimensional electrophoresis (IEF followed by SDS-PAGE) the presence of basic and acidic polypeptides has been demonstrated. The enzyme showed Km values of 5, 5.6 and 8 mu M for GTP, CTP and ATP, respectively, and the activity was inhibited by low levels of oc-amanitin and antibodies raised against bovine RNA polymerase II. By Western blot analysis the enzyme was found to cross-react with antibodies to bovine RNA polymerase II. RNA polymerase II from G. utilis is a phosphoprotein, the subunits RPB1 and RPB10 were found to be phosphorylated. Analysis of carboxy-terminal domain indicated that it was functionally redundant at least in case of nonspecific transcription, implicating its role in other nuclear processes, such as promoter specific initiation or transcription activation or RNA processing.

Item Type: Journal Article
Publication: Biochemistry & Molecular Biology International
Publisher: Academic Press Aust
Additional Information: Copyright of this article belongs to Academic Press Aust.
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 02 Jun 2011 10:23
Last Modified: 02 Jun 2011 10:23
URI: http://eprints.iisc.ac.in/id/eprint/38156

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