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  • Canonical nucleators are dispensable for stress granule assembly in Drosophila intestinal progenitors.

Canonical nucleators are dispensable for stress granule assembly in Drosophila intestinal progenitors.

Journal of cell science (2020-04-09)
Kasun Buddika, Ishara S Ariyapala, Mary A Hazuga, Derek Riffert, Nicholas S Sokol
要旨

Stressed cells downregulate translation initiation and assemble membrane-less foci termed stress granules (SGs). Although SGs have been extensively characterized in cultured cells, the existence of such structures in stressed adult stem cell pools remains poorly characterized. Here, we report that the Drosophila orthologs of the mammalian SG components AGO1, ATX2, CAPRIN, eIF4E, FMRP, G3BP, LIN-28, PABP and TIAR are enriched in adult fly intestinal progenitor cells, where they accumulate in small cytoplasmic messenger ribonucleoprotein complexes (mRNPs). Treatment with sodium arsenite or rapamycin reorganized these mRNPs into large cytoplasmic granules. Formation of these intestinal progenitor stress granules (IPSGs) depended on polysome disassembly, led to translational downregulation and was reversible. Although the canonical SG nucleators ATX2 and G3BP were sufficient for IPSG formation in the absence of stress, neither of them, nor TIAR, either individually or collectively, were required for stress-induced IPSG formation. This work therefore finds that IPSGs do not assemble via a canonical mechanism, raising the possibility that other stem cell populations employ a similar stress-response mechanism.

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ピューロマイシン 二塩酸塩 Streptomyces alboniger由来, powder, BioReagent, suitable for cell culture
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モノクローナル抗FLAG® M2抗体 マウス宿主抗体, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
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Immobilon®-P PVDF Membrane, 1 roll, 27 cm x 3.75 m, 0.45 µm pore size, Hydrophobic PVDF Transfer Membrane for western blotting.
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シクロヘキシミド, from microbial, ≥94% (TLC)
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シュナイダー昆虫培地, With L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for insect cell culture