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Roles of the Pumilio domain protein PUF3 in Trypanosoma brucei growth and differentiation

Published online by Cambridge University Press:  09 June 2020

K. Kamanyi Marucha  and
C. Clayton Open the ORCID record for C. Clayton [Opens in a new window]
K. Kamanyi Marucha
Affiliation:
Heidelberg University Centre for Molecular Biology (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
C. Clayton*
Affiliation:
Heidelberg University Centre for Molecular Biology (ZMBH), Im Neuenheimer Feld 282, D-69120 Heidelberg, Germany
*
Author for correspondence: C. Clayton, E-mail: cclayton@zmbh.uni-heidelberg.de
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Abstract

Trypanosomes strongly rely on post-transcriptional mechanisms to control gene expression. Several Opisthokont Pumilio domain proteins are known to suppress expression when bound to mRNAs. The Trypanosoma brucei Pumilio domain protein PUF3 is a cytosolic mRNA-binding protein that suppresses expression when tethered to a reporter mRNA. RNA-binding studies showed that PUF3 preferentially binds to mRNAs with a classical Pumilio-domain recognition motif, UGUA[U/C]AUU. RNA-interference-mediated reduction of PUF3 in bloodstream forms caused a minor growth defect, but the transcriptome was not affected. Depletion of PUF3 also slightly delayed differentiation to the procyclic form. However, both PUF3 genes could be deleted in cultured bloodstream- and procyclic-form trypanosomes. Procyclic forms without PUF3 also grew somewhat slower than wild-type, but ectopic expression of C-terminally tagged PUF3 impaired their viability. PUF3 was not required for RBP10-induced differentiation of procyclic forms to bloodstream forms. Mass spectrometry revealed no PUF3 binding partners that might explain its suppressive activity. We conclude that PUF3 may have a role in fine-tuning gene expression. Since PUF3 is conserved in all Kinetoplastids, including those that do not infect vertebrates, we suggest that it might confer advantages within the invertebrate host.

Keywords

mRNA decayPumiliotranslationTrypanosoma brucei
Type
Research Article
Information
Parasitology , Volume 147 , Issue 11 , September 2020 , pp. 1171 - 1183
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Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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Footnotes

*

Current address: Kisii University Medical Biochemistry, P.O. Box 408, 40200 Kisii, Kenya.

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