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PMID: 2100996 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The subunit structure of Saccharomyces cerevisiae transcription factor IIIC probed with a novel photocrosslinking reagent.

The EMBO journal ·Vol. 9 ·No. 7 ·1990-07-00 ·Pages 2197-205

Bartholomew B, Kassavetis GA, Braun BR, Geiduschek EP

Abstract

A photocrosslinking nucleotide, 5-[N-(p-azidobenzoyl)-3-aminoallyl]-deoxyuridine monophosphate (N3Rd-UMP), has been used to identify four polypeptides that are associated with the large Saccharomyces cerevisiae RNA polymerase III transcription factor TFIIIC, and to map the locations of these subunits along DNA when TFIIIC binds to the S.cerevisiae SUP4 tRNA(Tyr) gene. The 145 kd subunit of TFIIIC is primarily accessible to photocrosslinking from the vicinity of the box B + internal promoter element; 95 and 55 kd subunits are located on opposite sides of the DNA helix in the vicinity of the box A internal promoter element; a 135 kd subunit is less strongly crosslinked to the box A region and to a DNA segment between boxes B and A. DNA probes containing more than one N3RdUMP residue can form crosslinks between polypeptide chains. The specific circumstances of formation and the apparent mol. wts of two of these products lead to the tentative suggestion that a protomer of TFIIIC may contain two 95 kd subunits.

MeSH Terms
Azides/metabolism Base Sequence Binding, Competitive Cross-Linking Reagents/metabolism DNA Probes Kinetics Macromolecular Substances Molecular Sequence Data Molecular Weight Saccharomyces cerevisiae/genetics,metabolism Transcription Factors/genetics,metabolism,radiation effects Transcription Factors, TFIII Ultraviolet Rays Uracil Nucleotides/metabolism Uridine Monophosphate/analogs & derivatives,metabolism
Chemicals
Azides Cross-Linking Reagents DNA Probes Macromolecular Substances Transcription Factors Transcription Factors, TFIII Uracil Nucleotides transcription factor TFIIIC 5-(N-(4-azidobenzoyl)-3-aminoallyl)deoxyuridine 5'-monophosphate Uridine Monophosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Bartholomew B
Department of Biology and Center for Molecular Genetics, University of California, San Diego, La Jolla 92093.
Kassavetis G A
Braun B R
Geiduschek E P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1990-07-00
Pages
2197-205
Language
English
Region
England
NLM ID
8208664
PMCID
PMC551943
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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