Abstract
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.a 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 27C and after 1 hour at 39°C, and the mRNA content of 41°C heat shocks granules. We found that 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. Seventy mRNAs moved towards the polysomal fraction after the 39°C heat shock; surprisingly, many of these 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
Author summary When trypanosomes are inside tsetse flies, they have to cope with temperature variations from below 20°C up to nearly 40°C, due to diurnal variations and periodic intake of warm blood. The procyclic forms, which usually multiply in the midgut, are routinely cultured at 27°C in the laboratory. When they are heated to temperatures of 37°C and above, they shut down protein production, and at 41°C, mRNAs aggregate into granules. We show here that quite a large number of mRNAs are not included in granules and continue to be used for making proteins. Some of the proteins that continue to be made are needed in order to defend the cells against the effects of heat shock. Interestingly, however, a moderate heat shock stimulates expression of genes needed for the parasites to develop further into forms that can colonise the salivary glands. It thus seems possible that in the field, temperature variations might influence the efficiency with which of trypanosomes in tsetse flies become infective for mammals.