Identification of a domain within human TAFI48, a subunit of Selectivity Factor 1, that interacts with helix 2 of TBP

Shuping Xu, Roderick Hori

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

RNA polymerase I transcription in human cells requires Selectivity Factor 1, a multisubunit complex composed of the TATA-box-binding protein (TBP) and three TBP-associated factors (TAFs) called TAFI48, TAFI63 and TAFI110. Each of the Selectivity Factor 1 subunits binds directly to the other three components, but these interactions have not been characterized. This study is the initial identification and analysis of a TBP-binding domain within a Selectivity Factor 1 TAF. The interaction between human TBP and human TAFI48 was initially examined using the yeast two-hybrid assay, and a TBP-binding domain was identified in the carboxyl-terminus of human (h)TAFI48. Consistent with this result, the hTAFI48 carboxyl-terminus was able to bind directly to TBP in protein-protein interaction assays. When mutations were introduced into the hTAFI48 carboxyl-terminus, we identified changes in uncharged and positive residues that affect its interaction with TBP. By examining TBP mutants, residues within and adjacent to helix 2 of TBP, previously demonstrated to interact with subunits of other TBP-containing complexes [Transcription Factor IID (TFIID) and TFIIIB] were also found to diminish its affinity for the carboxyl-terminus of hTAFI48. The regions of hTAFI48 and TBP that interact are compared to those identified within other complexes containing TBP.

Original languageEnglish (US)
Pages (from-to)177-186
Number of pages10
JournalGene
Volume338
Issue number2
DOIs
StatePublished - Sep 1 2004

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TATA-Box Binding Protein
TATA-Binding Protein Associated Factors
Protein Binding
Transcription Factor TFIIIB
Transcription Factor TFIID
RNA Polymerase I
Two-Hybrid System Techniques
Proteins

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Identification of a domain within human TAFI48, a subunit of Selectivity Factor 1, that interacts with helix 2 of TBP. / Xu, Shuping; Hori, Roderick.

In: Gene, Vol. 338, No. 2, 01.09.2004, p. 177-186.

Research output: Contribution to journalArticle

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abstract = "RNA polymerase I transcription in human cells requires Selectivity Factor 1, a multisubunit complex composed of the TATA-box-binding protein (TBP) and three TBP-associated factors (TAFs) called TAFI48, TAFI63 and TAFI110. Each of the Selectivity Factor 1 subunits binds directly to the other three components, but these interactions have not been characterized. This study is the initial identification and analysis of a TBP-binding domain within a Selectivity Factor 1 TAF. The interaction between human TBP and human TAFI48 was initially examined using the yeast two-hybrid assay, and a TBP-binding domain was identified in the carboxyl-terminus of human (h)TAFI48. Consistent with this result, the hTAFI48 carboxyl-terminus was able to bind directly to TBP in protein-protein interaction assays. When mutations were introduced into the hTAFI48 carboxyl-terminus, we identified changes in uncharged and positive residues that affect its interaction with TBP. By examining TBP mutants, residues within and adjacent to helix 2 of TBP, previously demonstrated to interact with subunits of other TBP-containing complexes [Transcription Factor IID (TFIID) and TFIIIB] were also found to diminish its affinity for the carboxyl-terminus of hTAFI48. The regions of hTAFI48 and TBP that interact are compared to those identified within other complexes containing TBP.",
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