Cell cycle localization dynamics of mitochondrial DNA polymerase IC in African trypanosomes.

TitleCell cycle localization dynamics of mitochondrial DNA polymerase IC in African trypanosomes.
Publication TypeJournal Article
Year of Publication2018
AuthorsConcepciĆ³n-Acevedo J, Miller JC, Boucher MJ, Klingbeil MM
JournalMol Biol Cell
Volume29
Issue21
Pagination2540-2552
Date Published10/2018
ISSN1939-4586
KeywordsCell Cycle, DNA Replication, DNA, Kinetoplast, DNA-Directed DNA Polymerase, Gene Knockdown Techniques, Gene Silencing, Mitochondria, Protozoan Proteins, S Phase, Trypanosoma brucei brucei
Abstract

Trypanosoma brucei has a unique catenated mitochondrial DNA (mtDNA) network called kinetoplast DNA (kDNA). Replication of kDNA occurs once per cell cycle in near synchrony with nuclear S phase and requires the coordination of many proteins. Among these are three essential DNA polymerases (TbPOLIB, IC, and ID). Localization dynamics of these proteins with respect to kDNA replication stages and how they coordinate their functions during replication are not well understood. We previously demonstrated that TbPOLID undergoes dynamic localization changes that are coupled to kDNA replication events. Here, we report the localization of TbPOLIC, a second essential DNA polymerase, and demonstrate the accumulation of TbPOLIC foci at active kDNA replication sites (antipodal sites) during stage II of the kDNA duplication cycle. While TbPOLIC was undetectable by immunofluorescence during other cell cycle stages, steady-state protein levels measured by Western blot remained constant. TbPOLIC foci colocalized with the fraction of TbPOLID that localized to the antipodal sites. However, the partial colocalization of the two essential DNA polymerases suggests a highly dynamic environment at the antipodal sites to coordinate the trafficking of replication proteins during kDNA synthesis. These data indicate that cell cycle-dependent localization is a major regulatory mechanism for essential mtDNA polymerases during kDNA replication.

DOI10.1091/mbc.E18-02-0127
Alternate JournalMol. Biol. Cell
PubMed ID30133333
PubMed Central IDPMC6254582
Grant ListR01 AI066279 / AI / NIAID NIH HHS / United States