Identification of a transcription factor IIIA-interacting protein

Rodney J. Moreland, Michael E. Dresser, Justin S. Rodgers, Bruce A. Roe, Joan W. Conaway, Ronald C. Conaway, Jay S. Hanas

Research output: Contribution to journalArticlepeer-review

49 Scopus citations


Transcription factor IIIA (TFIIIA) activates 5S ribosomal RNA gene transcription in eukaryotes. The protein from vertebrates has nine contiguous Cys2His2 zinc fingers which function in nucleic acid binding, and a C-terminal region involved in transcription activation. In order to identify protein partners for TFIIIA, yeast two-hybrid screens were performed using the C-terminal region of Xenopus TFIIIA as an attractor and a rat cDNA library as a source of potential partners. A cDNA clone was identified which produced a protein in yeast that interacted with Xenopus TFIIIA but not with yeast TFIIIA. This rat clone was sequenced and the primary structure of the human homolog (termed TFIIIA-intP for TFIIIA-interacting protein) was determined from expressed sequence tags. In vitro interaction of recombinant human TFIIIA-intP with recombinant Xenopus TFIIIA was demonstrated by immunoprecipitation of the complex using anti-TFIIIA-intP antibody. Interaction of rat TFIIIA with rat TFIIIA-intP was indicated by co-chromatography of the two proteins on DEAE-5PW following fractionation of a rat liver extract on cation, anion and gel filtration resins. In a HeLa cell nuclear extract, recombinant TFIIIA-intP was able to stimulate TFIIIA-dependent transcription of the Xenopus SS ribosomal RNA gene but not TFIIIA-independent transcription of the human adenovirus VA RNA gene.

Original languageEnglish (US)
Pages (from-to)1986-1993
Number of pages8
JournalNucleic acids research
Issue number9
StatePublished - May 1 2000
Externally publishedYes

ASJC Scopus subject areas

  • Genetics


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