title: Biolayer Interferometry: A Novel Method to Elucidate Protein-Protein and Protein-DNA Interactions in the Mitochondrial DNA Replisome.
abstract: A lack of effective treatment for mitochondrial diseases prompts scientists to investigate the molecular processes that underlie their development. The major cause of mitochondrial diseases is dysfunction of the sole mitochondrial DNA polymerase, DNA polymerase γ (Pol γ). The development of treatment strategies will require a detailed characterization of the molecular properties of Pol γ. A novel technique, biolayer interferometry, allows one to monitor molecular interactions in real time, thus providing an insight into the kinetics of the process. Here, we present an application of the biolayer interferometry technique to characterize the fundamental reactions that Pol γ undergoes during the initiation phase of mitochondrial DNA replication: holoenzyme formation and binding to the primer-template.
authors.affiliation: Institute of Biosciences and Medical Technology, University of Tampere, Tampere, 33014, Finland.
authors.forename: Grzegorz L
authors.initials: GL
authors.lastname: Ciesielski
authors.affiliation: Institute of Biosciences and Medical Technology, University of Tampere, Tampere, 33014, Finland.|Fimlab Laboratories, Biokatu 4, Tampere, 33101, Finland.
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
authors.affiliation: Institute of Biosciences and Medical Technology, University of Tampere, Tampere, 33014, Finland. lskaguni@msu.edu.|Department of Biochemistry and Molecular Biology, Center for Mitochondrial Science and Medicine, Michigan State University, East Lansing, MI, 48824, USA. lskaguni@msu.edu.
authors.forename: Laurie S
authors.initials: LS
authors.lastname: Kaguni
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D000074002:DNA Polymerase gamma; D004261:DNA Replication / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D006801:Humans; D007368:Interferometry / Q000379:methods*; D008928:Mitochondria / Q000235:genetics; D028361:Mitochondrial Diseases / Q000235:genetics; D009097:Multienzyme Complexes / Q000235:genetics*; D000099272:Replisomes
chemical_list: D000074002:DNA Polymerase gamma; D004272:DNA, Mitochondrial; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D009097:Multienzyme Complexes; D000099272:Replisomes
keywords: Biolayer interferometry; Mitochondria; Mitochondrial replisome; Protein–protein interactions