Reference: Steffan JS, et al. (1996) The role of TBP in rDNA transcription by RNA polymerase I in Saccharomyces cerevisiae: TBP is required for upstream activation factor-dependent recruitment of core factor. Genes Dev 10(20):2551-63

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Abstract


Transcription of Saccharomyces cerevisiae rDNA by RNA polymerase I involves at least two transcription factors characterized previously: upstream activation factor (UAF) consisting of Rrn5p, Rrn9p, Rrn10p, and two more uncharacterized proteins; and core factor (CF) consisting of Rrn6p, Rrn7p, and Rrn11p. UAF interacts directly with an upstream element of the promoter and mediates its stimulatory function, and CF subsequently joins a stable preinitiation complex. The TATA-binding protein (TBP) has been known to be involved in transcription by all three nuclear RNA polymerases. We found that TBP interacts specifically with both UAF and CF, the interaction with UAF being stronger than that with CF. Using extracts from a TBP (I143N) mutant, it was shown that TBP is required for stimulation of transcription mediated by the upstream element, but not for basal transcription directed by a template without the upstream element. By template competition experiments, it was shown that TBP is required for UAF-dependent recruitment of CF to the rDNA promoter, explaining the TBP requirement for stimulatory activity of the upstream element. We also studied protein-protein interactions and found specific interactions of TBP with Rrn6p and with Rrn9p both in vitro and in the yeast two-hybrid system in vivo. Thus, these two interactions may be involved in the interactions of TBP with CF and UAF, respectively, contributing to the recruitment of CF to the rDNA promoter. Additionally, we observed an interaction between Rrn9p and Rrn7p both in vitro and in the two-hybrid system; thus, this interaction might also contribute to the recruitment of CF.

Reference Type
Journal Article | Research Support, U.S. Gov't, P.H.S.
Authors
Steffan JS, Keys DA, Dodd JA, Nomura M
Primary Lit For
RRN10 | RRN11 | SPT15 | RRN9 | RRN5 | RNA polymerase I upstream activating factor complex | RNA polymerase I core factor complex | DNA-directed RNA Polymerase I complex
Additional Lit For
RRN7 | RRN6

Gene Ontology Annotations 18 entries for 1 gene


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Gene/ComplexQualifierGene Ontology TermAnnotation ExtensionEvidenceSourceAssigned On
SPT15enablesRNA polymerase I general transcription initiation factor bindingIDASGD2013-08-07
SPT15involved inRNA polymerase I preinitiation complex assemblyIMPSGD2020-01-31
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN6IPI with RRN11SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN7IPI with RRN11SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN11IPI with RRN11SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN9IPI with RRN11SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN6IPI with RRN9SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN7IPI with RRN9SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN11IPI with RRN9SGD2013-08-07
SPT15enablesRNA polymerase I general transcription initiation factor bindinghas input RRN9IPI with RRN9SGD2013-08-07
Showing 1 to 10 of 18 entries

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 7 entries for 4 genes

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InteractorInteractorAssayAnnotationActionModification
RRN6SPT15Reconstituted Complexmanually curatedHit-BaitNo Modification
RRN6SPT15Two-hybridmanually curatedHit-BaitNo Modification
RRN7RRN9Reconstituted Complexmanually curatedHit-BaitNo Modification
RRN7RRN9Two-hybridmanually curatedHit-BaitNo Modification
SPT15RRN9Reconstituted Complexmanually curatedBait-HitNo Modification
SPT15RRN9Two-hybridmanually curatedHit-BaitNo Modification
SPT15RRN9Two-hybridmanually curatedBait-HitNo Modification
Showing 1 to 7 of 7 entries