Unraveling ATAD3

Presenter

Katelyn Belmore

Campus

UMass Amherst

Sponsor

Elizabeth Vierling, Department of Biochemistry and Molecular Biology, UMass Amherst

Schedule

Session 4, 2:30 PM - 3:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A33, Campus Center Auditorium, Row 2 (A21-A40) [Poster Location Map]

Abstract

The primary objective of this research is to better understand the molecular function of the mitochondrial membrane-embedded ATPase family AAA Domain-containing Protein 3 (ATAD3) in Arabidopsis thaliana. AAA+ proteins, characterized by conserved motifs within their AAA+ domains, operate as ATPases, serving as regulatory components that modulate cellular processes through controlled ATP hydrolysis. In addition, the topology of ATAD3, spanning from the mitochondrial matrix to the cytosol, supports a role in inter-organellar communication, which may extend to vital cellular processes beyond the mitochondrion. ATAD3 proteins are highly conserved essential proteins found in essentially all eukaryotes except the Fungi. Mutation of these proteins in humans leads to severe growth defects and lethality. The consequences of ATAD3 disruption in A. thaliana reveal slow growth and alterations in mitochondrial DNA organization and oxidative phosphorylation, while deletion of ATAD3s is lethal. To study the specific molecular function of ATAD3 we aim to identify potential substrates of the ATPase domain, which has homology to molecular chaperones that act to modify protein structure. For this purpose, “TRAP” mutations were introduced into the AAA+ domain to prevent ATP hydrolysis, which would act to prevent substrate release, along with an affinity “SPOT” tag to allow purification of the modified ATAD3 protein. The current goal is to obtain homozygous plants carrying the TRAP mutant genes. Recovery of ATAD3-TRAP proteins followed by mass spectrometry should allow identification of potential ATAD3 substrates, providing new insight into ATAD3 function. We are also creating new null mutant alleles of ATAD3 proteins using CRISPR technology.

Keywords

Plant Biology, Biochemistry, Mitochondria, Climate Change

Research Area

Biological Organisms

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