Autoinhibition of TFIIIB70 binding by the tetratricopeptide repeat-containing subunit of TFIIIC

Robyn D. Moir, Karen V. Puglia, Ian M. Willis

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

An important step in the assembly of RNA polymerase (pol) III transcription complexes on tRNA and 5 S genes is the interaction between the tetratricopeptide repeat (TPR)-containing subunit of TFIIIC (TFIIIC131) and the TFIIB-related subunit of TFIIIB (TFIIIB70/Brf1). A fragment of TFIIIC131 that contains the hydrophilic amino terminus and two TPR arrays, with five and four repeats, respectively (Nt-TPR9), is sufficient to support an interaction with TFIIIB70. Here we evaluate the contribution of each TPR array to TFIIIB70 binding. Both TPR arrays bind independently to TFIIIB70 with TPR6-9 having a 4-fold higher apparent affinity than TPR1-5. However, the TPR arrays are not sufficient for a high affinity interaction with TFIIIB70. The addition of amino-terminal sequences increases the affinity of TPR1-5 18-fold to create a high affinity TFIIIB70 binding site (Nt-TPR5, 44 ± 6 nM). Although the Nt-TPR5 and TPR6-9 fragments are contained entirely within the Nt-TPR9 fragment, the affinity of the latter is significantly lower than either of these smaller fragments. The results demonstrate that the TFIIIB70 binding sites in TFIIIC131 are subject to autoinhibition. We propose that the binding of TFIIIB70 to these sites within the TFIIIC complex may proceed in an ordered fashion.

Original languageEnglish (US)
Pages (from-to)694-701
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number1
DOIs
StatePublished - Jan 4 2002

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Transcription Factor TFIIIB
Transcription Factor TFIIB
Binding Sites
RNA Polymerase III
Transcription
Transfer RNA
Genes
transcription factor TFIIIC

ASJC Scopus subject areas

  • Biochemistry

Cite this

Autoinhibition of TFIIIB70 binding by the tetratricopeptide repeat-containing subunit of TFIIIC. / Moir, Robyn D.; Puglia, Karen V.; Willis, Ian M.

In: Journal of Biological Chemistry, Vol. 277, No. 1, 04.01.2002, p. 694-701.

Research output: Contribution to journalArticle

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