Abstract
We observed that heat shock of Caenorhabditis elegans leads to the formation of nuclear double-stranded RNA (dsRNA) foci, detectable with a dsRNA-specific monoclonal antibody. These foci significantly overlap with nuclear HSF-1 granules. To investigate the molecular mechanism(s) underlying dsRNA foci formation, we used RNA-seq to globally characterize total RNA and immunoprecipitated dsRNA from control and heat shocked worms. We find a subset of both sense and antisense transcripts enriched in the dsRNA pool by heat shock overlap with dsRNA transcripts enriched by deletion of tdp-1, which encodes the C. elegans ortholog of TDP-43. Interestingly, transcripts involved in translation are over-represented in the dsRNAs induced by either heat shock or deletion of tdp-1. Also enriched in the dsRNA transcripts are sequences downstream of annotated genes (DoGs), which we globally quantified with a new algorithm. To validate these observations, we used fluorescence in situ hybridization (FISH) to confirm both antisense and downstream of gene transcription for eif-3.B, one of the affected loci we identified.
Highlights
Cytoplasmic proteotoxic stress induced by temperatures outside of the optimal range for cells or organisms triggers the heat shock response (HSR) [1]
Using the J2 antibody for immunohistochemistry, we found J2 double-stranded RNA (dsRNA) foci in nuclear regions that partially overlapped with nuclear HSF-1 stress granules when drSI-13 worms were heat shocked for 35 ̊C for 40 minutes (Fig 1A–1C)
We show that heat shock induces nuclear dsRNA foci that partially overlap with HSF-1 nuclear stress granules
Summary
Cytoplasmic proteotoxic stress induced by temperatures outside of the optimal range for cells or organisms triggers the heat shock response (HSR) [1]. The response to heat shock is multifaceted and regulation of both transcription and translation occurs. Transcriptional responses include formation of stress granules, alternative splicing, and aberrant transcriptional termination [2,3,4,5]. The HSR is a highly conserved transcriptional response and is driven largely by the heat shock transcription factor HSF1 [6]. HSF1 is a monomer in the cytoplasm and nucleus. HSF1 undergoes homotrimerization and binds to DNA heat shock elements (HSE) and initiates the transcription of heat shock protein genes [7,8].
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