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RNA pol II subunit RPB7 is required for RNA pol I-mediated transcription in Trypanosoma brucei.

ABSTRACT: In the protozoan parasite Trypanosoma brucei, the two main surface glycoprotein genes are transcribed by RNA polymerase I (pol I) instead of RNA pol II, the polymerase committed to the production of mRNA in eukaryotes. This unusual feature might be accomplished by the recruitment of specific subunits or cofactors that allow pol I to transcribe protein-coding RNAs. Here, we report that transcription mediated by pol I requires TbRPB7, a dissociable subunit of the pol II complex. TbRPB7 was found to interact with two pol I-specific subunits, TbRPA1 and TbRPB6z. Pol I-specific transcription was affected on depletion of TbRPB7 in run-on assays, whereas recombinant TbRPB7 increased transcription driven by a pol I promoter. These results represent a unique example of a functional RNA polymerase chimaera consisting of a core pol I complex that recruits a specific pol II subunit.

SUBMITTER: Penate X

PROVIDER: S-EPMC2658562 | biostudies-literature | 2009 Mar

REPOSITORIES: biostudies-literature

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RNA pol II subunit RPB7 is required for RNA pol I-mediated transcription in Trypanosoma brucei.

Peñate Xenia X López-Farfán Diana D Landeira David D Wentland Amy A Vidal Isabel I Navarro Miguel M

EMBO reports 20090123 3

In the protozoan parasite Trypanosoma brucei, the two main surface glycoprotein genes are transcribed by RNA polymerase I (pol I) instead of RNA pol II, the polymerase committed to the production of mRNA in eukaryotes. This unusual feature might be accomplished by the recruitment of specific subunits or cofactors that allow pol I to transcribe protein-coding RNAs. Here, we report that transcription mediated by pol I requires TbRPB7, a dissociable subunit of the pol II complex. TbRPB7 was found t ...[more]

PMID: 19165144

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Dynein Light Chain LC8 Is Required for RNA Polymerase I-Mediated Transcription in Trypanosoma brucei, Facilitating Assembly and Promoter Binding of Class I Transcription Factor A.

Project description:Dynein light chain LC8 is highly conserved among eukaryotes and has both dynein-dependent and dynein-independent functions. Interestingly, LC8 was identified as a subunit of the class I transcription factor A (CITFA), which is essential for transcription by RNA polymerase I (Pol I) in the parasite Trypanosoma brucei. Given that LC8 has never been identified with a basal transcription factor and that T. brucei relies on RNA Pol I for expressing the variant surface glycoprotein (VSG), the key protein in antigenic variation, we investigated the CITFA-specific role of LC8. Depletion of LC8 from mammalian-infective bloodstream trypanosomes affected cell cycle progression, reduced the abundances of rRNA and VSG mRNA, and resulted in rapid cell death. Sedimentation analysis, coimmunoprecipitation of recombinant proteins, and bioinformatic analysis revealed an LC8 binding site near the N terminus of the subunit CITFA2. Mutation of this site prevented the formation of a CITFA2-LC8 heterotetramer and, in vivo, was lethal, affecting assembly of a functional CITFA complex. Gel shift assays and UV cross-linking experiments identified CITFA2 as a promoter-binding CITFA subunit. Accordingly, silencing of LC8 or CITFA2 resulted in a loss of CITFA from RNA Pol I promoters. Hence, we discovered an LC8 interaction that, unprecedentedly, has a basal function in transcription.

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