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PMID: 10393904 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A protein-protein interaction map of yeast RNA polymerase III.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 96 ·No. 14 ·1999-07-06 ·Pages 7815-20

Flores A, Briand JF, Gadal O, Andrau JC, Rubbi L, Van Mullem V, Boschiero C, Goussot M, Marck C, Carles C, Thuriaux P, Sentenac A, Werner M

Abstract

The structure of the yeast RNA polymerase (pol) III was investigated by exhaustive two-hybrid screening using a library of random genomic fragments fused to the Gal4 activation domain. This procedure allowed us to identify contacts between individual polypeptides, localize the contact domains, and deduce a protein-protein interaction map of the multisubunit enzyme. In all but one case, pol III subunits were able to interact in vivo with one or sometimes two partner subunits of the enzyme or with subunits of TFIIIC. Four subunits that are common to pol I, II, and III (ABC27, ABC14.5, ABC10alpha, and ABC10beta), two that are common to pol I and III (AC40 and AC19), and one pol III-specific subunit (C11) can associate with defined regions of the two large subunits. These regions overlapped with highly conserved domains. C53, a pol III-specific subunit, interacted with a 37-kDa polypeptide that copurifies with the enzyme and therefore appears to be a unique pol III subunit (C37). Together with parallel interaction studies based on dosage-dependent suppression of conditional mutants, our data suggest a model of the pol III preinitiation complex.

MeSH Terms
Binding Sites Conserved Sequence Macromolecular Substances Open Reading Frames Peptide Library RNA Polymerase I/chemistry,metabolism RNA Polymerase II/chemistry,metabolism RNA Polymerase III/chemistry,genetics,metabolism Recombinant Fusion Proteins/chemistry Saccharomyces cerevisiae/enzymology,genetics Transcription Factors/chemistry,metabolism Transcription Factors, TFIII Transcription, Genetic
Chemicals
Macromolecular Substances Peptide Library Recombinant Fusion Proteins Transcription Factors Transcription Factors, TFIII transcription factor TFIIIC RNA Polymerase II RNA Polymerase I RNA Polymerase III
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Flores A
Service de Biochimie et Génétique Moléculaire, Bât. 142, Commissariat à l'Energie Atomique/Saclay, F-91191 Gif-sur-Yvette Cedex, France.
Briand J F
Gadal O
Andrau J C
Rubbi L
Van Mullem V
Boschiero C
Goussot M
Marck C
Carles C
Thuriaux P
Sentenac A
Werner M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.96.14.7815
Published
1999-07-06
Pages
7815-20
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC22144
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.96.14.7815 PMC Full Text View journal details
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