A protein–protein interaction map of yeast RNA polymerase III
- 6 July 1999
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 96 (14) , 7815-7820
- https://doi.org/10.1073/pnas.96.14.7815
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.Keywords
This publication has 55 references indexed in Scilit:
- Two large subunits of the fission yeast RNA polymerase II provide platforms for the assembly of small subunitsJournal of Molecular Biology, 1998
- Two conformations of RNA polymerase II revealed by electron crystallographyJournal of Molecular Biology, 1997
- Toward a functional analysis of the yeast genome through exhaustive two-hybrid screensNature Genetics, 1997
- Mapping the Contacts of Yeast TFIIIB and RNA Polymerase III at Various Distances from the Major Groove of DNA by DNA Photoaffinity LabelingPublished by Elsevier ,1996
- Molecular assembly of RNA polymerase II from the fission yeast Schizosaccharomyces pombe: subunit–subunit contact network involving Rpb5Genes to Cells, 1996
- The C-terminal Domain Revealed in the Structure of RNA Polymerase IIJournal of Molecular Biology, 1996
- Studies on the transformation of intact yeast cells by the LiAc/SS‐DNA/PEG procedureYeast, 1995
- Encounters of Saccharomyces cererisiae RNA Polymerase III with its Transcription Factors during RNA Chain ElongationJournal of Molecular Biology, 1994
- A yeast TFIIB-related factor involved in RNA polymerase III transcription.Genes & Development, 1992
- Three-dimensional structure of Escherichia coli RNA polymerase holoenzyme determined by electron crystallographyNature, 1989