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- W2894544169 abstract "Improper maintenance of the mitochondrial genome progressively disrupts cellular respiration and causes severe metabolic disorders commonly termed mitochondrial diseases. Mitochondrial single-stranded DNA binding protein (mtSSB) is an essential component of the mtDNA replication machinery. We utilized single-molecule methods to examine the modes by which human mtSSB binds DNA to help define protein interactions at the mtDNA replication fork. Direct visualization of individual mtSSB molecules by atomic force microscopy (AFM) revealed a random distribution of mtSSB tetramers bound to extended regions of single-stranded DNA (ssDNA), strongly suggesting non-cooperative binding by mtSSB. Selective binding to ssDNA was confirmed by AFM imaging of individual mtSSB tetramers bound to gapped plasmid DNA substrates bearing defined single-stranded regions. Shortening of the contour length of gapped DNA upon binding mtSSB was attributed to DNA wrapping around mtSSB. Tracing the DNA path in mtSSB–ssDNA complexes with Dual-Resonance-frequency-Enhanced Electrostatic force Microscopy established a predominant binding mode with one DNA strand winding once around each mtSSB tetramer at physiological salt conditions. Single-molecule imaging suggests mtSSB may not saturate or fully protect single-stranded replication intermediates during mtDNA synthesis, leaving the mitochondrial genome vulnerable to chemical mutagenesis, deletions driven by primer relocation or other actions consistent with clinically observed deletion biases." @default.
- W2894544169 created "2018-10-05" @default.
- W2894544169 creator A5013164620 @default.
- W2894544169 creator A5018238846 @default.
- W2894544169 creator A5025498462 @default.
- W2894544169 creator A5051576642 @default.
- W2894544169 creator A5058936762 @default.
- W2894544169 date "2018-09-26" @default.
- W2894544169 modified "2023-10-15" @default.
- W2894544169 title "Single-molecule DREEM imaging reveals DNA wrapping around human mitochondrial single-stranded DNA binding protein" @default.
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- W2894544169 doi "https://doi.org/10.1093/nar/gky875" @default.
- W2894544169 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6265486" @default.