Presentation Title

Factors that Affect the TDP-43 Aggregation During Stress Conditions

Faculty Mentor

Brian M Zid

Start Date

17-11-2018 12:30 PM

End Date

17-11-2018 2:30 PM

Location

HARBESON 14

Session

POSTER 2

Type of Presentation

Poster

Subject Area

biological_agricultural_sciences

Abstract

TAR DNA binding protein 43 (TDP-43) is a nuclear protein that contains two RNA recognition motifs (RRMs) and a Glycine-rich region in its C-terminal domain. The function of TDP-43 is unclear, but it has been found to aggregate in deposits observed in patients with Amniotic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD). Recently it has been demonstrated that TDP-43 colocalizes with Stress Granules (SG), RNP granules that form during stress conditions by the aggregation of proteins and RNAs. Our research group is interested in understanding the aggregation behavior of TDP-43 by varying protein concentration, as well as altering environmental conditions. To understand the aggregation of TDP-43 we used yeast as a model: wild type human TDP-43 protein attached to the mRuby2 fluorescent protein marker is driven by the Tet-On promoter which is dependent on doxycycline concentration. Based on the results obtained from concentration dependency of TDP-43, we determined that the optimal concentration is 40 ng/uL of doxycycline. Based on the optimal concentration test, we followed TDP-43 over time and observed that the expression of TDP-43 increased over time and predominantly localized to the nucleus. Surprisingly we found that when our cultures were infected with another fungus this caused TDP-43 to translocate from the nucleus to the cytoplasm and form cytoplasmic aggregates. To determine the colocalization of the aggregates in our yeast model we will use nuclear fluorescent markers as well as other RNP granule markers. Overall, we hope to learn more about the factors that control TDP-43 aggregation.

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Nov 17th, 12:30 PM Nov 17th, 2:30 PM

Factors that Affect the TDP-43 Aggregation During Stress Conditions

HARBESON 14

TAR DNA binding protein 43 (TDP-43) is a nuclear protein that contains two RNA recognition motifs (RRMs) and a Glycine-rich region in its C-terminal domain. The function of TDP-43 is unclear, but it has been found to aggregate in deposits observed in patients with Amniotic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD). Recently it has been demonstrated that TDP-43 colocalizes with Stress Granules (SG), RNP granules that form during stress conditions by the aggregation of proteins and RNAs. Our research group is interested in understanding the aggregation behavior of TDP-43 by varying protein concentration, as well as altering environmental conditions. To understand the aggregation of TDP-43 we used yeast as a model: wild type human TDP-43 protein attached to the mRuby2 fluorescent protein marker is driven by the Tet-On promoter which is dependent on doxycycline concentration. Based on the results obtained from concentration dependency of TDP-43, we determined that the optimal concentration is 40 ng/uL of doxycycline. Based on the optimal concentration test, we followed TDP-43 over time and observed that the expression of TDP-43 increased over time and predominantly localized to the nucleus. Surprisingly we found that when our cultures were infected with another fungus this caused TDP-43 to translocate from the nucleus to the cytoplasm and form cytoplasmic aggregates. To determine the colocalization of the aggregates in our yeast model we will use nuclear fluorescent markers as well as other RNP granule markers. Overall, we hope to learn more about the factors that control TDP-43 aggregation.