Investigation of Physical Interactions between Essential Mitochondrial DNA Polymerase Paralogs in Trypanosoma brucei

Presenter

Henry Meinke Raughley

Campus

UMass Amherst

Sponsor

Michele M. Klingbeil, Department of Microbiology, UMass Amherst

Schedule

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

Location

Poster Board A42, Campus Center Auditorium, Row 3 (A41-A60) [Poster Location Map]

Abstract

African sleeping sickness is a fatal vector-borne disease if left untreated. Trypanosoma brucei, the causative agent, contains unique mitochondrial DNA, that is a network of catenated minicircle and maxicircle DNA molecules called kinetoplast DNA (kDNA). Replication of the kDNA network is an essential precisely timed cellular event, thus identifying kDNA replication proteins as potential drug targets. Coordination of many proteins including three independently essential DNA polymerases (POLIB, POLIC, and POLID) is required for network replication. During replication POLIC and POLID exhibit a dynamic localization pattern colocalizing with minicircle replication progeny at two antipodal sites (APS) that flank the kDNA network.
Interestingly, RNA interference (RNAi) of POLID results in progressive loss of minicircle progeny, but also a decrease in POLIC APS localization during kDNA replication and decreased POLIC protein levels. We hypothesize that disruption of POLIC APS localization results from physical interactions with the other essential DNA polymerases. To investigate whether depletion of POLIB protein also impacts POLIC APS localization, I will evaluate POLIC localization, protein levels, and impact on minicircle progeny during POLIB RNAi. To more directly test physical interactions, I will immunoprecipitate POLIC from cell lines containing epitope-tagged versions of POLIB or POLID. Physical interactions between or among the essential DNA polymerases could serve as targets for small molecule inhibitors.

Keywords

DNA replication, Parasitology, Molecular Biology, Neglected Tropical Disease

Research Area

Biological Organisms

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