Studying mtDNA replication using in vitro reconstituted systems | 34792
Journal of Proteomics & Bioinformatics

Journal of Proteomics & Bioinformatics
Open Access

ISSN: 0974-276X

Studying mtDNA replication using in vitro reconstituted systems

7th International Conference on Proteomics & Bioinformatics

October 24-26, 2016 Rome, Italy

Jay P Uhler

University of Gothenburg, Sweden

Posters & Accepted Abstracts: J Proteomics Bioinform

Abstract :

Mitochondria are essential for the production of cellular energy and contain their own genomes (mtDNA) that encode 13 subunits of the respiratory chain, 22 tRNAs and 2 rRNAs. The remaining >1500 mitochondrial proteins are nuclear encoded including those required for mtDNA maintenance. The human mitochondrial genome is an approximately 16.5kb circular molecule; mutations in which can lead to mitochondrial disease. mtDNA is replicated by a dedicated mitochondrial replication machinery that includes the replicative DNA polymerase POL�?³, the TWINKLE helicase and mitochondrial single stranded DNA binding protein. However, many other DNA replication factors remain to be identified and studied. Moreover, the regulation of mtDNA replication initiation, elongation and termination are not yet fully understood. To help address these unresolved questions, our research group reconstituted a minimal mitochondrial replisome in vitro over 10 years ago. This in vitro mtDNA replication system is based on purified recombinant proteins and specifically designed DNA templates. Based on this approach, many basic mechanisms have been addressed such as leading/lagging strand replication and RNA priming/ removal. The system also enables in vitro modeling of mitochondrial diseases by using disease-based mutant proteins or by reconstituting aberrant replication processes. Most recently, the system has been useful in performing screens to uncover the effects of various drug compounds on mtDNA replication.

Biography :