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Molecular and Cellular Biology, November 2004, p. 9610-9618, Vol. 24, No. 21
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.21.9610-9618.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Functional Characterization of a Trypanosoma brucei TATA-Binding Protein-Related Factor Points to a Universal Regulator of Transcription in Trypanosomes

Jia-peng Ruan,1 George K. Arhin,2 Elisabetta Ullu,2,3 and Christian Tschudi1,2*

Departments of Epidemiology and Public Health,1 Internal Medicine,2 Cell Biology, Yale University Medical School, New Haven, Connecticut3

Received 16 May 2004/ Returned for modification 19 July 2004/ Accepted 13 August 2004

Transcriptional mechanisms remain poorly understood in trypanosomatid protozoa. In particular, there is no knowledge about the function of basal transcription factors, and there is an apparent rarity of promoters for protein-coding genes transcribed by RNA polymerase (Pol) II. Here we describe a Trypanosoma brucei factor related to the TATA-binding protein (TBP). Although this TBP-related factor (TBP-related factor 4 [TRF4]) has about 31% identity to the TBP core domain, several key residues involved in TATA box binding are not conserved. Depletion of the T. brucei TRF4 (TbTRF4) by RNA interference revealed an essential role in RNA Pol I, II, and III transcription. Using chromatin immunoprecipitation, we further showed that TRF4 is recruited to the Pol I-transcribed procyclic acidic repetitive genes, Pol II-transcribed spliced leader RNA genes, and Pol III-transcribed U-snRNA and 7SL RNA genes, thus supporting a role for TbTRF4 in transcription performed by all three nuclear RNA polymerases. Finally, a search for TRF4 binding sites in the T. brucei genome led to the identification of such sites in the 3' portion of certain protein-coding genes, indicating a unique aspect of Pol II transcription in these organisms.

* Corresponding author. Mailing address: Department of Epidemiology and Public Health, Yale University Medical School, 295 Congress Ave., New Haven, CT 06536-0812. Phone: (203) 785-7332. Fax: (203) 785-7329. E-mail: christian.tschudi{at}yale.edu.

Molecular and Cellular Biology, November 2004, p. 9610-9618, Vol. 24, No. 21
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.21.9610-9618.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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