Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids

0449774 - FGÚ 2016 RIV GR eng J - Journal Article
Olejár, Tomáš - Pajuelo-Reguera, David - Alán, Lukáš - Dlasková, Andrea - Ježek, Petr
Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids.
Molecular Medicine Reports. Roč. 12, č. 4 (2015), s. 5185-5190. ISSN 1791-2997. E-ISSN 1791-3004
R&D Projects: GA ČR(CZ) GAP302/10/0346; GA MŠMT(CZ) EE2.3.30.0025
Institutional support: RVO:67985823
Keywords : mitochondrial nucleoid * single-stranded DNA-binding protein * photoconvertible fluorescent protein Eos
Subject RIV: EA - Cell Biology
Impact factor: 1.559, year: 2015

A small limited number of large aggregates of mtSSB EosFP tagged protein started to form in the cells, which exhibited a great co localization with the noted coupled positions. Antibody staining and 3D immunocytochemistry confirmed that nucleoid components such as TFAM and DNA were co localized with these aggregates. Furthermore, the observed reduction of the mtDNA copy number in mtSSB EosFP transfected cells suggested a possible impairment of nucleoid function. In conclusion, the present study demonstrated that coupled nucleoids are synchronized by mtSSB EosFP overexpression and visualized through their equal binding capacity to mtSSB EosFP tagged protein. This observation suggested parallel replication and transcription activity of nucleoid couples native from a parental one. Preserved coupling in late stages of artificial EosFP mediated aggregation of tagged proteins suggested a rational manner of mitochondrial branching that may be cell-type specifically dependent on hierarchical nucleoid replication
Permanent Link: http://hdl.handle.net/11104/0251977