Glycolysis modulates trypanosome glycoprotein expression as revealed by an RNAi library
Open Access
- 2 September 2002
- journal article
- research article
- Published by Springer Nature in The EMBO Journal
- Vol. 21 (17) , 4429-4438
- https://doi.org/10.1093/emboj/cdf474
Abstract
RNA interference (RNAi) is a powerful tool for identifying gene function in Trypanosoma brucei . We generated an RNAi library, the first of its kind in any organism, by ligation of genomic fragments into the vector pZJMβ. After transfection at ∼5‐fold genome coverage, trypanosomes were induced to express double‐stranded RNA and screened for reduced con canavalin A (conA) binding. Since this lectin binds the surface glycoprotein EP‐procyclin, we predicted that cells would lose affinity to conA if RNAi silenced genes affecting EP‐procyclin expression or modification. We found a cell line in which RNAi switches expression from glycosylated EP‐procyclins to the unglycosylated GPEET‐procyclin. This switch results from silencing a hexokinase gene. The relationship between procyclin expression and glycolysis was supported by silencing other genes in the glycolytic pathway, and confirmed by observation of a similar upregulation of GPEET‐ procyclin when parental cells were grown in medium depleted of glucose. These data suggest that T.brucei ‘senses’ changes in glucose level and modulates procyclin expression accordingly.Keywords
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