title: TbISWI regulates multiple polymerase I (Pol I)-transcribed loci and is present at Pol II transcription boundaries in Trypanosoma brucei.
abstract: The unicellular eukaryote Trypanosoma brucei is unusual in having very little transcriptional control. The bulk of the T. brucei genome is constitutively transcribed by RNA polymerase II (Pol II) as extensive polycistronic transcription units. Exceptions to this rule include several RNA Pol I transcription units such as the VSG expression sites (ESs), which are mono-allelically expressed. TbISWI, a member of the SWI2/SNF2 related chromatin remodeling ATPases, plays a role in repression of Pol I-transcribed ESs in both bloodstream- and procyclic-form T. brucei. We show that TbISWI binds both active and silent ESs but is depleted from the ES promoters themselves. TbISWI knockdown results in an increase in VSG transcripts from the silent VSG ESs. In addition to its role in the repression of the silent ESs, TbISWI also contributes to the downregulation of the Pol I-transcribed procyclin loci, as well as nontranscribed VSG basic copy arrays and minichromosomes. We also show that TbISWI is enriched at a number of strand switch regions which form the boundaries between Pol II transcription units. These strand switch regions are the presumed sites of Pol II transcription initiation and termination and are enriched in modified histones and histone variants. Our results indicate that TbISWI is a versatile chromatin remodeler that regulates transcription at multiple Pol I loci and is particularly abundant at many Pol II transcription boundaries in T. brucei.
authors.affiliation: Division of Cell and Molecular Biology, Sir Alexander Fleming Building, Imperial College London-South Kensington, Imperial College Road, London, United Kingdom.
authors.forename: Tara M
authors.initials: TM
authors.lastname: Stanne
authors.forename: Manish
authors.initials: M
authors.lastname: Kushwaha
authors.forename: Matthew
authors.initials: M
authors.lastname: Wand
authors.forename: Jesse E
authors.initials: JE
authors.lastname: Taylor
authors.forename: Gloria
authors.initials: G
authors.lastname: Rudenko
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism*; D000483:Alleles; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins; D005434:Flow Cytometry; D015966:Gene Expression Regulation, Fungal; D006657:Histones; D012318:RNA Polymerase I / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014346:Trypanosoma brucei brucei / Q000201:enzymology / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D006657:Histones; C086468:ISWI protein; D014157:Transcription Factors; D012319:RNA Polymerase II; D012318:RNA Polymerase I; D000251:Adenosine Triphosphatases