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

Archana Sareen; Priya Choudhry; Surbhi Mehta; Nimisha Sharma

doi:10.1016/j.bbrc.2005.05.015

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

Archana Sareen, Priya Choudhry, Surbhi Mehta, Nimisha Sharma

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	763-770
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	332
Issue number	3
DOIs	https://doi.org/10.1016/j.bbrc.2005.05.015
State	Published - Jul 8 2005
Externally published	Yes

Bibliographical note

Funding Information:
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.

Keywords

RNA polymerase II
Rpb4
Rpb7

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10.1016/j.bbrc.2005.05.015

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title = "Mapping the interaction site of Rpb4 and Rpb7 subunits of RNA polymerase II in Saccharomyces cerevisiae",

abstract = "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.",

keywords = "RNA polymerase II, Rpb4, Rpb7",

author = "Archana Sareen and Priya Choudhry and Surbhi Mehta and Nimisha Sharma",

note = "Funding Information: 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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AU - Choudhry, Priya

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AU - Sharma, Nimisha

N1 - Funding Information: 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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N2 - 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.

AB - 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.

KW - RNA polymerase II

KW - Rpb4

KW - Rpb7

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

Abstract

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Keywords

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