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Expression of hsrω-RNAi transgene prior to heat shock specifically compromises accumulation of heat shock-induced Hsp70 in Drosophila melanogaster

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Cell Stress and Chaperones Aims and scope

An Erratum to this article was published on 20 October 2015

Abstract

A delayed organismic lethality was reported in Drosophila following heat shock when developmentally active and stress-inducible noncoding hsrω-n transcripts were down-regulated during heat shock through hs-GAL4-driven expression of the hsrω-RNAi transgene, despite the characteristic elevation of all heat shock proteins (Hsp), including Hsp70. Here, we show that hsrω-RNAi transgene expression prior to heat shock singularly prevents accumulation of Hsp70 in all larval tissues without affecting transcriptional induction of hsp70 genes and stability of their transcripts. Absence of the stress-induced Hsp70 accumulation was not due to higher levels of Hsc70 in hsrω-RNAi transgene-expressing tissues. Inhibition of proteasomal activity during heat shock restored high levels of the induced Hsp70, suggesting very rapid degradation of the Hsp70 even during the stress when hsrω-RNAi transgene was expressed ahead of heat shock. Unexpectedly, while complete absence of hsrω transcripts in hsrω 66 homozygotes (hsrω-null) did not prevent high accumulation of heat shock-induced Hsp70, hsrω-RNAi transgene expression in hsrω-null background blocked Hsp70 accumulation. Nonspecific RNAi transgene expression did not affect Hsp70 induction. These observations reveal that, under certain conditions, the stress-induced Hsp70 can be selectively and rapidly targeted for proteasomal degradation even during heat shock. In the present case, the selective degradation of Hsp70 does not appear to be due to down-regulation of the hsrω-n transcripts per se; rather, this may be an indirect effect of the expression of hsrω-RNAi transgene whose RNA products may titrate away some RNA-binding proteins which may also be essential for stability of the induced Hsp70.

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Acknowledgments

We thank Dr. K. V. Prasanth (USA) for H5 antibody, Dr. R. M. Tanguay (Canada) for anti-Hsp90 antibody, and Dr. M. B. Evgen’ev (Russia) for 7Fb and 7.10.3 antibodies. Bloomington Stock Centre is acknowledged for providing various stocks used in present study. This work was supported by the CEIB-II grant from Department of Biotechnology, Govt. of India, and by the Board of Research in Nuclear Sciences (Department of Atomic Energy, Govt. of India) through Raja Ramanna Fellowship to SCL. We thank the Department of Science & Technology, Govt. of India (New Delhi), and the Banaras Hindu University for Confocal Microscopy facility in our laboratory. AKS has been supported as Senior Research Fellow by the Council of Scientific & Industrial Research (New Delhi) and as Research Associate by the Department of Biotechnology, Govt. of India.

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  1. Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, 221005, India

    Anand K. Singh & Subhash C. Lakhotia

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Correspondence to Subhash C. Lakhotia.

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Singh, A.K., Lakhotia, S.C. Expression of hsrω-RNAi transgene prior to heat shock specifically compromises accumulation of heat shock-induced Hsp70 in Drosophila melanogaster . Cell Stress and Chaperones 21, 105–120 (2016). https://doi.org/10.1007/s12192-015-0644-6

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