Trypanosoma brucei variant surface glycoprotein regulation involves coupled activation/inactivation and chromatin remodeling of expression sites.
Trypanosoma brucei is an extracellular protozoan parasite that cycles between mammalian hosts and the tsetse vector. In bloodstream-form trypanosomes, only one variant surface glycoprotein gene (VSG) expression site (ES) is active at any time. Transcriptional switching between ESs results in antigenic variation. No VSG is transcribed in the insect procyclic stage. We have used bacteriophage T7 RNA polymerase (T7RNAP) to study the transcriptional accessibility of ES chromatin in vivo. We show that T7RNAP-mediated transcription from chromosomally integrated T7 promoters is repressed along the entire length of the ES in the procyclic form, but not in the bloodstream form, suggesting that the accessible chromatin of inactive bloodstream-form ESs is remodeled upon differentiation to yield a structure that is no longer permissive for T7RNAP-mediated transcription. In the bloodstream form, replacing the active ES promoter with a T7 promoter, which is incapable of sustaining high-level transcription of the entire ES, prompts an ES switch. These data suggest two distinct mechanisms for ES regulation: a chromatin-mediated developmental silencing of the ES in the procyclic form and a rapid coupled mechanism for ES activation and inactivation in the bloodstream form.
Navarro
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M
Laboratory of Molecular Parasitology, The Rockefeller University (Box 185), 1230 York Avenue, New York, NY 10021-6399, USA.
Cross
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Wirtz
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D000818:Animals; D001483:Base Sequence; D002843:Chromatin / Q000378:metabolism*; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014346:Trypanosoma brucei brucei / Q000378:metabolism*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000235:genetics*; D014764:Viral Proteins
D002843:Chromatin; D017931:DNA Primers; D014643:Variant Surface Glycoproteins, Trypanosoma; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases