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HSF-1 is a Regulator of miRNA Expression in Caenorhabditis elegans
EP26900
Poster Title: HSF-1 is a Regulator of miRNA Expression in Caenorhabditis elegans
Submitted on 08 Feb 2018
Author(s): Alana Snyder; Jessica Brunquell, Ph.D.; Feng Cheng, Ph.D.; Sandy Westerheide, Ph.D.
Affiliations: USF Morsani College of Medicine
This poster was presented at USF Health Research Day 2018
Poster Views: 518
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Poster Information
Abstract: Objectives
To discover the role of HSF-1 in regulating and coordinating stress-dependent and stress-independent processes through post-transcriptional regulation of miRNAs in Caenorhabditis elegans.

Methods
Age synchronous wild-type C. elegans were grown at 23 ˚C under RNA interference conditions. These were carried out on standard plates supplemented with 50 µg/mL ampicillin and 1 mM isopropyl-beta-D-thiogalactopyranoside seeded with E. coli HT115 bacteria containing an empty vector or sequence-verified hsf-1 RNAi. Heat shock treatment was carried out at the L4 stage by submerging plates in a 33 ˚C water bath for 30 minutes. miRNA was isolated from biological duplicates using TRIzol, then cleaned and sequenced. miRDeep2 software was used to identify known miRNAs from the miRNA-sequencing data and mapped to a C. elegans reference genome. The fold-change of each miRNA was obtained by normalizing the corresponding miRNA for each treatment condition to the control. Validation was done by qRT-PCR. mirWIP software and TargetScan were used to determine the target predictions for the miRNA-mRNA interactions.

Results
HSF-1 alters global miRNA abundance during and independently of heat shock, suggesting HSF-1 as a regulator of miRNA expression. Because miRNA normally has inhibitory effects on mRNA, the miRNAs that are upregulated by HSF-1 during heat shock, are responsible for suppressing the correlated biological processes, including those involved in cell fate commitment, signal transduction, protein kinase activity, signaling cascades, and development. Several miRNAs are normally downregulated during HS by HSF-1 and result in increased activation of mRNAs related to cuticle growth, reproduction, and cell signaling. Independently of heat shock, HSF-1 was found to downregulated miRNAs involved in inducing the processes of signaling, ion binding, and development. Several other miRNAs were normally upregulated independently of heat shock, including those involved in suppressing growth, development, cytoskeletal organization, epithelial development, and cuticle development. Significance in fold-changes was determined by the Benjamini-Hochberg correction for multiple testing.

Conclusions
The miRNA-sequencing performed in this experiment and the data obtained show a possible role of HSF-1 post-transcriptionally regulating heat stress-dependent and -independent processes in C. elegans. Some of the implicated processes are longevity, development, and metabolism.
Summary: The transcription factor HSF-1 was found to regulate development, metabolism, and longevity through control of microRNA expression in C. elegans.Report abuse »
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