Abstract
Kinetoplastids rely heavily on post-transcriptional mechanisms for control of gene expression, with regulation of mRNA processing, translation and degradation by RNA-binding proteins. ZC3H32 is a cytosolic mRNA-binding protein with three non-canonical CCCH zinc finger domains. It is much more abundant in bloodstream-form Trypanosoma brucei than in procyclic forms. Tethering of ZC3H32 to a reporter mRNA suppressed translation and resulted in mRNA degradation, and deletion analysis suggested that this activity was present in both the N- and C-terminal domains, but not the central zinc finger-containing domain. Tandem affinity purification, however, revealed no interaction partners that might account for this activity. RNASeq analyses did not yield any evidence for sequence-specific binding or regulation of specific mRNAs. The presence of ZC3H32 homologues in monogenetic and free-living Euglenids also argues against a role in developmental regulation, although its function may have diverged in evolution. T. brucei ZC3H32 might be implicated in basal mRNA metabolism, with this role perhaps being taken over by another protein in procyclic forms.
Highlights
Organisms in the Phylum Euglenozoa rely on trans splicing to process mRNAs from polycistronically transcribed precursors
This has been most extensively analysed in the parasitic trypanosomes and leishmanias, in which it has been shown that regulation of individual mRNAs relies heavily on post-transcriptional mechanisms [1, 2]
ZC3H32 homologues are present in all Trypanasomatida genomes sequenced to date, including all salivarian trypanosomes, Leishmania, Endotrypanum, and Crithidia and the more distantly-related Angomonas and Bodo saltans, a free-living bacteriovore
Summary
Organisms in the Phylum Euglenozoa rely on trans splicing to process mRNAs from polycistronically transcribed precursors This has been most extensively analysed in the parasitic trypanosomes and leishmanias, in which it has been shown that regulation of individual mRNAs relies heavily on post-transcriptional mechanisms [1, 2]. In Trypanosoma brucei, the level of an mRNA is determined by the number of gene copies, the decay rates in both nucleus and cytosol, and the processing efficiency [3]. The latter three processes are in turn strongly influenced by RNA-binding proteins [4,5,6]. ZC3H32 is about 20 times more abundant in bloodstream forms than procyclic forms, with five
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