Journal
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume 172, Issue 2, Pages 99-106Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.molbiopara.2010.03.016
Keywords
Coupling; RNA polymerase II; Transcription; trans splicing; Trypanosoma
Categories
Funding
- Deutsche Forschungsgemeinschaft
- Wellcome Trust
- Croucher Foundation
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Capping of mRNAs is strictly coupled to RNA polymerase II transcription and there is evidence, mainly from metazoans, that other steps in pre-mRNA processing show a similar linkage. In trypanosomes, however, the mRNA cap is supplied by a trans spliced leader sequence. Thus pre-mRNAs transcribed by RNA Polymerase I are capped by trans splicing, and translation-competent transgenic mRNAs can be produced by RNA Polymerase I and T7 RNA polymerase so long as the primary transcript has a splice acceptor signal. We quantified the efficiency of processing of trypanosome pre-mRNAs produced from a plasmid integrated either at the tubulin locus, or in an rRNA spacer, and transcribed by RNA polymerase II, RNA polymerase I or T7 RNA polymerase. The processing efficiencies were similar for primary transcripts from the tubulin locus, produced by RNA polymerase II, and for RNA from an rRNA spacer, transcribed by RNA polymerase I. Primary transcripts produced by 17 RNA polymerase from the tubulin locus were processed almost as well. There was therefore no evidence for recruitment of the 3'-splicing apparatus by the RNA polymerase. Abundant transcripts transcribed from the rRNA locus by T7 RNA polymerase were somewhat less efficiently processed. (C) 2010 Elsevier B.V. All rights reserved.
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