1. 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
- Author
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Clementine Merce, Christine Clayton, Monica Terrao, and Igor Minia
- Subjects
0301 basic medicine ,Hot Temperature ,Protozoan Proteins ,Gene Expression ,RNA-binding protein ,RNA-binding proteins ,Biochemistry ,Heat Shock Response ,Salivary Glands ,Translational regulation ,Medicine and Health Sciences ,Cellular Stress Responses ,lcsh:Public aspects of medicine ,Messenger RNA ,Granule (cell biology) ,Nucleic acids ,Infectious Diseases ,Cell Processes ,Cellular Structures and Organelles ,Anatomy ,Sequence Analysis ,RNA, Protozoan ,Research Article ,lcsh:Arctic medicine. Tropical medicine ,Tsetse Flies ,lcsh:RC955-962 ,Trypanosoma brucei brucei ,Biology ,Research and Analysis Methods ,03 medical and health sciences ,Stress granule ,Exocrine Glands ,Sequence Motif Analysis ,Polysome ,P-bodies ,Genetics ,Animals ,RNA, Messenger ,Heat shock ,Molecular Biology Techniques ,Sequencing Techniques ,Molecular Biology ,AU-rich element ,030102 biochemistry & molecular biology ,Biology and life sciences ,Public Health, Environmental and Occupational Health ,Proteins ,lcsh:RA1-1270 ,Cell Biology ,Chaperone Proteins ,030104 developmental biology ,Gene Expression Regulation ,Polyribosomes ,RNA ,Protein Translation ,Ribosomes ,Digestive System ,Heat-Shock Response - 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°Cā41°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., Author Summary 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. In the laboratory, procyclic forms (the form that multiplies in the midgut), are routinely cultured at 27°C. When procyclic forms are heated to temperatures of 37°C and above, they decrease 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.
- Published
- 2016