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Mapping the interaction site of Rpb4 and Rpb7 subunits of RNA polymerase II in Saccharomyces cerevisiae

Sareen, Archana and Choudhry, Priya and Mehta, Surbhi and Sharma, Nimisha (2005) Mapping the interaction site of Rpb4 and Rpb7 subunits of RNA polymerase II in Saccharomyces cerevisiae. Biochemical and Biophysical Research Communications, 332 (3). pp. 763-770. ISSN 0006-291X. doi:10.1016/j.bbrc.2005.05.015.

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Rpb4 and Rpb7, the fourth and the seventh largest subunits of RNA polymerase II, form a heterodimer in Saccharomyces cerevisiae. To identify the site of interaction between these subunits, we constructed truncation mutants of both these proteins and carried out yeast two hybrid analysis. Deletions in the amino and carboxyl terminal domains of Rpb7 abolished its interaction with Rpb4. In comparison, deletion of up to 49 N-terminal amino acids of Rpb4 reduced its interaction with Rpb7. Complete abolishment of interaction between Rpb4 and Rpb7 occurred by truncation of 1-106, 1-142, 108-221, 172-221 or 198-221 amino acids of Rpb4. Use of the yeast two-hybrid analysis in conjunction with computational analysis of the recently reported crystal structure of Rpb4/Rpb7 sub-complex allowed us to identify regions previously not suspected to be involved in the functional interaction of these proteins. Taken together, our results have identified the regions that are involved in interaction between the Rpb4 and Rpb7 subunits of S. cerevisiae RNA polymerase II in vivo.

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Additional Information:Copyright © 2005 Elsevier. Received 25 April 2005. Available online 13 May 2005. This study was supported by a grant from University Grants Commission. We are very grateful to Dr. Erica Golemis (FCCC, Philadelphia, Pennsylvania) for the yeast two hybrid strain and vectors. We thank Dr. Debasis Mohanty for his invaluable help with the bioinformatics analysis and use of the bioinformatics facility at National Institute of Immunology is acknowledged. We thank Dr. N. Raghuram and Dr. Jyoti Jaiswal for helpful comments on the manuscript. We also thank Dr. Rajnish Kaushik, Dr. Shibani Mitra-Kaushik, and Veenu Aishwarya, and the past and present members of the laboratory for all their help and suggestions.
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University Grants Commission (India)UNSPECIFIED
Subject Keywords:RNA polymerase II; Rpb4; Rpb7
Issue or Number:3
Record Number:CaltechAUTHORS:20140416-133159466
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:44986
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Deposited On:21 Apr 2014 23:09
Last Modified:10 Nov 2021 16:59

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