Session 1E: Exploring Whether the RID Effector Domain in the MARTX Toxin of Vibrio cholerae Is Sufficient to Downregulate the ERK Pathway

Session Number

Session 1E: 4th Presentation

Advisor(s)

Karla J.F. Satchell, Northwestern University

Location

Room A113

Start Date

28-4-2017 8:30 AM

End Date

28-4-2017 9:45 AM

Abstract

Vibrio cholerae is a gram negative bacterium that is spread through the fecal-oral route that can cause cholera when infected. It is commonly found in third world countries in contaminated water supplies. This study specifically looks into the Rho-Inactivation Domain (RID) effector domain found in the Multifunctional Autoprocessing Repeats-In-Toxin (MARTX) toxin found inside of the El Tor strain of this bacterium. The purpose of investigating RID is to see if it is sufficient enough to inhibit the Extracellular Signal–regulated Kinases(ERK) pathway, which may inhibit cytokines. To study this, HeLa cells were intoxicated with RID via a lethal factor/protective antigen delivery mechanism. Using these intoxications, western blots were performed with an antibody that detected the “on” form of ERK1/2 and another antibody that detected the “on and off” forms of ERK1/2. The western blots showed that RID is insufficient to downregulate ERK. The reason behind this can be due to insufficient amounts of ERK present in our samples. Therefore, future directions for this study involve activating ERK to increase the amount of ERK present.

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Apr 28th, 8:30 AM Apr 28th, 9:45 AM

Session 1E: Exploring Whether the RID Effector Domain in the MARTX Toxin of Vibrio cholerae Is Sufficient to Downregulate the ERK Pathway

Room A113

Vibrio cholerae is a gram negative bacterium that is spread through the fecal-oral route that can cause cholera when infected. It is commonly found in third world countries in contaminated water supplies. This study specifically looks into the Rho-Inactivation Domain (RID) effector domain found in the Multifunctional Autoprocessing Repeats-In-Toxin (MARTX) toxin found inside of the El Tor strain of this bacterium. The purpose of investigating RID is to see if it is sufficient enough to inhibit the Extracellular Signal–regulated Kinases(ERK) pathway, which may inhibit cytokines. To study this, HeLa cells were intoxicated with RID via a lethal factor/protective antigen delivery mechanism. Using these intoxications, western blots were performed with an antibody that detected the “on” form of ERK1/2 and another antibody that detected the “on and off” forms of ERK1/2. The western blots showed that RID is insufficient to downregulate ERK. The reason behind this can be due to insufficient amounts of ERK present in our samples. Therefore, future directions for this study involve activating ERK to increase the amount of ERK present.