期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 290, 期 45, 页码 26954-26967出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M115.679019
关键词
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资金
- Wellcome Trust
- BBSRC [BB/J01477X/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/J01477X/1] Funding Source: researchfish
Background: Eukaryotes typically encode a range of ISWI chromatin remodeling complexes with different functions. Results: We have identified and analyzed three novel ISWI partners in the early branching eukaryote Trypanosoma brucei. Conclusion:T. brucei appears to have a single major ISWI complex. Significance: This unusually simple ISWI configuration could be a consequence of the relative lack of transcriptional regulation in this ancient eukaryote. ISWI chromatin remodelers are highly conserved in eukaryotes and are important for the assembly and spacing of nucleosomes, thereby controlling transcription initiation and elongation. ISWI is typically associated with different subunits, forming specialized complexes with discrete functions. In the unicellular parasite Trypanosoma brucei, which causes African sleeping sickness, TbISWI down-regulates RNA polymerase I (Pol I)-transcribed variant surface glycoprotein (VSG) gene expression sites (ESs), which are monoallelically expressed. Here, we use tandem affinity purification to determine the interacting partners of TbISWI. We identify three proteins that do not show significant homology with known ISWI-associated partners. Surprisingly, one of these is nucleoplasmin-like protein (NLP), which we had previously shown to play a role in ES control. In addition, we identify two novel ISWI partners, regulator of chromosome condensation 1-like protein (RCCP) and phenylalanine/tyrosine-rich protein (FYRP), both containing protein motifs typically found on chromatin proteins. Knockdown of RCCP or FYRP in bloodstream form T. brucei results in derepression of silent variant surface glycoprotein ESs, as had previously been shown for TbISWI and NLP. All four proteins are expressed and interact with each other in both major life cycle stages and show similar distributions at Pol I-transcribed loci. They are also found at Pol II strand switch regions as determined with ChIP. ISWI, NLP, RCCP, and FYRP therefore appear to form a single major ISWI complex in T. brucei (TbIC). This reduced complexity of ISWI regulation and the presence of novel ISWI partners highlights the early divergence of trypanosomes in evolution.
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