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The role of genomic location and flanking 3′UTR in the generation of functional levels of variant surface glycoprotein in Trypanosoma brucei

Ridewood, Sophie, Ooi, Cher-Pheng, Hall, Belinda, Trenaman, Anna, Wand, Nadina Vasileva, Sioutas, Georgios, Scherwitzl, Iris and Rudenko, Gloria (2017) The role of genomic location and flanking 3′UTR in the generation of functional levels of variant surface glycoprotein in Trypanosoma brucei Molecular Microbiology, 106 (4). pp. 614-634.

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Trypanosoma brucei faces relentless immune attack in the mammalian bloodstream, where it is protected by an essential coat of Variant Surface Glycoprotein (VSG) comprising ∼10% total protein. The active VSG gene is in a Pol I‐transcribed telomeric expression site (ES). We investigated factors mediating these extremely high levels of VSG expression by inserting ectopic VSG117 into VSG221 expressing T. brucei. Mutational analysis of the ectopic VSG 3′UTR demonstrated the essentiality of a conserved 16‐mer for mRNA stability. Expressing ectopic VSG117 from different genomic locations showed that functional VSG levels could be produced from a gene 60 kb upstream of its normal telomeric location. High, but very heterogeneous levels of VSG117 were obtained from the Pol I‐transcribed rDNA. Blocking VSG synthesis normally triggers a precise precytokinesis cell‐cycle checkpoint. VSG117 expression from the rDNA was not adequate for functional complementation, and the stalled cells arrested prior to cytokinesis. However, VSG levels were not consistently low enough to trigger a characteristic ‘VSG synthesis block’ cell‐cycle checkpoint, as some cells reinitiated S phase. This demonstrates the essentiality of a Pol I‐transcribed ES, as well as conserved VSG 3′UTR 16‐mer sequences for the generation of functional levels of VSG expression in bloodstream form T. brucei.

Item Type: Article
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine
Authors :
Ridewood, Sophie
Ooi, Cher-Pheng
Trenaman, Anna
Wand, Nadina Vasileva
Sioutas, Georgios
Scherwitzl, Iris
Rudenko, Gloria
Date : 14 November 2017
Funders : Wellcome Trust
DOI : 10.1111/mmi.13838
Copyright Disclaimer : © 2017 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Depositing User : Clive Harris
Date Deposited : 12 Sep 2019 14:46
Last Modified : 12 Sep 2019 14:46

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