Translation Regulation and RNA Granule Formation after Heat Shock of Procyclic Form Trypanosoma brucei: Many Heat-Induced mRNAs Are also Increased during Differentiation to Mammalian-Infective Forms.

Igor Minia,Clementine Merce,Christine Clayton,Monica Terrao,Alvaro Acosta-Serrano

doi:10.1371/journal.pntd.0004982

Igor Minia, Clementine Merce + Show 3 more

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https://doi.org/10.1371/journal.pntd.0004982

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Abstract

African trypanosome procyclic forms multiply in the midgut of tsetse flies, and are routinely cultured at 27°C. Heat shocks of 37°C and above result in general inhibition of translation, and severe heat shock (41°C) results in sequestration of mRNA in granules. The mRNAs that are bound by the zinc-finger protein ZC3H11, including those encoding refolding chaperones, escape heat-induced translation inhibition. At 27°C, ZC3H11 mRNA is predominantly present as an untranslated cytosolic messenger ribonucleoprotein particle, but after heat shocks of 37°C—41°C, the ZC3H11 mRNA moves into the polysomal fraction. To investigate the scope and specificities of heat-shock translational regulation and granule formation, we analysed the distributions of mRNAs on polysomes at 27°C and after 1 hour at 39°C, and the mRNA content of 41°C heat shock granules. We found that mRNAs that bind to ZC3H11 remained in polysomes at 39°C and were protected from sequestration in granules at 41°C. As previously seen for starvation stress granules, the mRNAs that encode ribosomal proteins were excluded from heat-shock granules. 70 mRNAs moved towards the polysomal fraction after the 39°C heat shock, and 260 increased in relative abundance. Surprisingly, many of these mRNAs are also increased when trypanosomes migrate to the tsetse salivary glands. It therefore seems possible that in the wild, temperature changes due to diurnal variations and periodic intake of warm blood might influence the efficiency with which procyclic forms develop into mammalian-infective forms.

Highlights

African trypanosomes, like all other organisms investigated so far, respond to heat shock by repressing general protein synthesis, while enhancing or retaining synthesis of proteins that are required to survive or recover from heat stress [1]
When trypanosomes are inside tsetse flies, they have to cope with temperature variations from below 20°C up to 37°C, due to diurnal variations and periodic intake of warm blood
We were interested in knowing which mRNAs show regulation similar to that of ZC3H11, since we hoped in that way to identify conserved sequence motifs

Summary

Introduction

Like all other organisms investigated so far, respond to heat shock by repressing general protein synthesis, while enhancing or retaining synthesis of proteins that are required to survive or recover from heat stress [1]. Trypanosomes lack the ability to control the transcription of individual protein-coding genes [2,3,4]. The selectivity of the heat shock response, like other changes in gene expression, relies on post-transcriptional mechanisms. Trypanosoma brucei procyclic forms are the forms that grow inside the tsetse fly midgut. In natural infections, these forms migrate to the proventriculus, developing into epimastigotes, and from there to the salivary glands where they become metacyclic forms which are infective for mammals [6]. The parasites differentiate into non-dividing short stumpy forms [7], which are pre-adapted for differentiation into procyclic forms upon uptake by tsetse [8]

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