Title

Regulation of GADD34 and CReP mRNA Expression in the Unfolded Protein Response

Poster Number

10C

Lead Author Major

Biological Sciences

Lead Author Status

Senior

Format

Poster Presentation

Faculty Mentor Name

Doug Weiser

Faculty Mentor Email

dweiser@pacific.edu

Faculty Mentor Department

Biological Sciences

Additional Faculty Mentor Name

Lisa Wrischnik

Additional Faculty Mentor Email

lwrischnik@pacific.edu

Additional Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

The failure to balance protein synthesis, folding, and degradation in the endoplasmic reticulum (ER) leads to the accumulation of unfolded proteins, leading to ER stress. Cells respond to ER stress by activating a stress response signaling pathway known as the Unfolded Protein Response (UPR). The UPR induces phosphorylation of eIF2α (Eukaryotic Initiation Factor 2) to attenuate global protein synthesis, allowing for a chance to clear misfolded proteins. This function is opposed by eIF2α phosphatases, which contain a catalytic subunit, Protein Phosphatase 1, and a scaffolding protein, either GADD34 or CReP. Inhibition of eIF2α phosphatases has shown to promote survival in cell types by prolonging the effects of the UPR. Despite the considerable clinical interest in eIF2α phosphatase inhibiting drugs, much is unknown about the mechanism of action of GADD34 and CReP. Zebrafish are an ideal model for this research because they are a good mimic of what happens in humans, and provide the ability to shut down or activate GADD34 and CReP in different tissues at different stages during ER stress and its recovery. Understanding the gene regulation of GADD34 and CReP can be done by observing the changes in gene expression in zebrafish embryos after the induction of ER stress. Primers for the genes of interest (BIP, CHOP, GADD34, and CReP) were designed to test for changes in levels of gene expression. The 24hpf (hours post fertilization) zebrafish embryos were treated with tunicamycin, thapsigargin, and salubrinal to induce ER stress and DMSO for a negative control for 4 and 24 hours. The RNA was purified from the treated embryos to perform quantitative PCR (qPCR) to look at changes in gene levels to understand when eIF2α phosphatases are active.

Location

DeRosa University Center Ballroom

Start Date

27-4-2018 10:00 AM

End Date

27-4-2018 12:00 PM

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Apr 27th, 10:00 AM Apr 27th, 12:00 PM

Regulation of GADD34 and CReP mRNA Expression in the Unfolded Protein Response

DeRosa University Center Ballroom

The failure to balance protein synthesis, folding, and degradation in the endoplasmic reticulum (ER) leads to the accumulation of unfolded proteins, leading to ER stress. Cells respond to ER stress by activating a stress response signaling pathway known as the Unfolded Protein Response (UPR). The UPR induces phosphorylation of eIF2α (Eukaryotic Initiation Factor 2) to attenuate global protein synthesis, allowing for a chance to clear misfolded proteins. This function is opposed by eIF2α phosphatases, which contain a catalytic subunit, Protein Phosphatase 1, and a scaffolding protein, either GADD34 or CReP. Inhibition of eIF2α phosphatases has shown to promote survival in cell types by prolonging the effects of the UPR. Despite the considerable clinical interest in eIF2α phosphatase inhibiting drugs, much is unknown about the mechanism of action of GADD34 and CReP. Zebrafish are an ideal model for this research because they are a good mimic of what happens in humans, and provide the ability to shut down or activate GADD34 and CReP in different tissues at different stages during ER stress and its recovery. Understanding the gene regulation of GADD34 and CReP can be done by observing the changes in gene expression in zebrafish embryos after the induction of ER stress. Primers for the genes of interest (BIP, CHOP, GADD34, and CReP) were designed to test for changes in levels of gene expression. The 24hpf (hours post fertilization) zebrafish embryos were treated with tunicamycin, thapsigargin, and salubrinal to induce ER stress and DMSO for a negative control for 4 and 24 hours. The RNA was purified from the treated embryos to perform quantitative PCR (qPCR) to look at changes in gene levels to understand when eIF2α phosphatases are active.