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PMID: 12796299 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

RNA polymerase I transcribes procyclin genes and variant surface glycoprotein gene expression sites in Trypanosoma brucei.

Eukaryotic cell ·Vol. 2 ·No. 3 ·2003-06-00 ·Pages 542-51

Günzl A, Bruderer T, Laufer G, Schimanski B, Tu LC, Chung HM, Lee PT, Lee MG

Abstract

In eukaryotes, RNA polymerase (pol) I exclusively transcribes the large rRNA gene unit (rDNA) and mRNA is synthesized by RNA pol II. The African trypanosome, Trypanosoma brucei, represents an exception to this rule. In this organism, transcription of genes encoding the variant surface glycoprotein (VSG) and the procyclins is resistant to alpha-amanitin, indicating that it is mediated by RNA pol I, while other protein-coding genes are transcribed by RNA pol II. To obtain firm proof for this concept, we generated a T. brucei cell line which exclusively expresses protein C epitope-tagged RNA pol I. Using an anti-protein C immunoaffinity matrix, we specifically depleted RNA pol I from transcriptionally active cell extracts. The depletion of RNA pol I impaired in vitro transcription initiated at the rDNA promoter, the GPEET procyclin gene promoter, and a VSG gene expression site promoter but did not affect transcription from the spliced leader (SL) RNA gene promoter. Fittingly, induction of RNA interference against the RNA pol I largest subunit in insect-form trypanosomes significantly reduced the relative transcriptional efficiency of rDNA, procyclin genes, and VSG expression sites in vivo whereas that of SL RNA, alphabeta-tubulin, and heat shock protein 70 genes was not affected. Our studies unequivocally show that T. brucei harbors a multifunctional RNA pol I which, in addition to transcribing rDNA, transcribes procyclin genes and VSG gene expression sites.

MeSH Terms
Animals DNA, Ribosomal/genetics Gene Expression Gene Expression Regulation Genes, Protozoan Membrane Glycoproteins/genetics,metabolism Promoter Regions, Genetic Protozoan Proteins/genetics,metabolism RNA Polymerase I/metabolism RNA, Protozoan/metabolism Transcription, Genetic Trypanosoma brucei brucei/genetics,metabolism Variant Surface Glycoproteins, Trypanosoma/genetics,metabolism
Chemicals
DNA, Ribosomal GPEET protein, Trypanosoma brucei Membrane Glycoproteins Protozoan Proteins RNA, Protozoan Variant Surface Glycoproteins, Trypanosoma procyclic acidic repetitive protein, Trypanosoma RNA Polymerase I
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Günzl Arthur
Medizinisch-Naturwissenschaftliches Forschungsinstitut der Universität Tübingen, 72074 Tübingen, Germany.
Bruderer Thomas
Laufer Gabriele
Schimanski Bernd
Tu Lan-Chun
Chung Hui-Min
Lee Pei-Tseng
Lee Mary Gwo-Shu
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Article Info
Journal
Eukaryotic cell
Abbr.
Eukaryot Cell
ISSN
1535-9778
DOI
10.1128/EC.2.3.542-551.2003
Published
2003-06-00
Pages
542-51
Language
English
Region
United States
NLM ID
101130731
PMCID
PMC161450
Subset
IM
Grants
NIAID NIH HHS · R01 AI028953 · United States
NIAID NIH HHS · AI28953 · United States
External Links
PubMed source DOI: 10.1128/EC.2.3.542-551.2003 PMC Full Text View journal details
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