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Volume 155, Issue 5

Mitochondrial nucleoids from the yeast : expansion of the repertoire of proteins associated with mitochondrial DNA Free

Abstract

Molecules of mitochondrial DNA (mtDNA) are packed into nucleic acid–protein complexes termed mitochondrial nucleoids (mt-nucleoids). In this study, we analysed mt-nucleoids of the yeast , which harbours a linear form of the mitochondrial genome. To identify conserved as well as specific features of mt-nucleoids in this species, we employed two strategies for analysis of their components. First, we investigated the protein composition of mt-nucleoids isolated from mitochondria, determined N-terminal amino acid sequences of 14 proteins associated with the mt-nucleoids and identified corresponding genes. Next, we complemented the list of mt-nucleoid components with additional candidates identified in the complete genome sequence of as homologues of mt-nucleoid proteins. Our approach revealed several known mt-nucleoid proteins as well as additional components that expand the repertoire of proteins associated with these cytological structures. In particular, we identified and purified the protein Gcf1, which is abundant in the mt-nucleoids and exhibits structural features in common with the mtDNA packaging protein Abf2 from . We demonstrate that Gcf1p co-localizes with mtDNA, has DNA-binding activity , and is able to stabilize mtDNA in the Δ mutant, all of which points to a role in the maintenance of the mitochondrial genome. Importantly, in contrast to Abf2p, analysis of Gcf1p predicted the presence of a coiled-coil domain and a single high-mobility group (HMG) box, suggesting that it represents a novel type of mitochondrial HMG protein.

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): DAPI, 4′,6-diamidino-2-phenylindole , EMSA, electrophoretic mobility shift assay , GST, glutathione S-transferase , HMG, high-mobility group , mt-nucleoid, mitochondrial nucleoid , mtDNA, mitochondrial DNA , mtHMG, mitochondrial HMG protein , mtTBP, mitochondrial telomere-binding protein , NP-40, Nonidet P40 and yEGFP3, yeast enhanced version 3 of GFP
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2009-05-01
2024-04-19
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Mitochondrial nucleoids from the yeast Candida parapsilosis: expansion of the repertoire of proteins associated with mitochondrial DNA
Microbiology 155, 1558 (2009); https://doi.org/10.1099/mic.0.027474-0
/content/journal/micro/10.1099/mic.0.027474-0
/content/journal/micro/10.1099/mic.0.027474-0
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