Journal Article

Analysis of Mitochondrial DNA Nucleoids in Wild-Type and a Mutant Strain of <i>Saccharomyces Cerevisiae</i> that Lacks the Mitochondrial HMG Box Protein Abf2p

Scott M. Newman, Olga Zelenaya-Troitskaya, Philip S. Perlman and Ronald A. Butow

in Nucleic Acids Research

Volume 24, issue 2, pages 386-389
Published in print January 1996 | ISSN: 0305-1048
Published online January 1996 | e-ISSN: 1362-4962 | DOI: https://dx.doi.org/10.1093/nar/24.2.386
Analysis of Mitochondrial DNA Nucleoids in Wild-Type and a Mutant Strain of Saccharomyces Cerevisiae that Lacks the Mitochondrial HMG Box Protein Abf2p

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DNA-protein complexes (nucleoids) are believed to be the segregating unit of mitochondrial DNA (mtDNA) in Saccharomyces cerevisiae. A mitochondrial HMG box protein, Abf2p, is needed for maintenance of mtDNA in cells grown on rich dextrose medium, but is dispensible in glycerol grown cells. As visualized by 4′,6′-diamino-2-phenylindole staining, mtDNA nucleoids in mutant cells lacking Abf2p (Δabf2) are diffuse compared with those in wild-type cells. We have isolated mtDNA nucleoids and characterized two mtDNA-protein complexes, termed NCLDp-2 and NCLDs-2, containing distinct but overlapping sets of polypeptides. This protocol yields similar nucleoid complexes from the Δabf2 mutant, although several proteins appear lacking from NCLDs-2. Segments of mtDNA detected with probes to COXII, VAR1 and ori5 sequences are equally sensitive to DNase I digestion in NCLDs-2 and NCLDp-2 from wild-type cells and from the Dabf2 mutant. However, COXII and VAR1 sequences are 4-to 5-fold more sensitive to DNase I digestion of mtDNA in toluene-permeabilized mitochondria from the Δabf2 mutant than from wild-type cells, but no difference in DNase I sensitivity was detected with the ori5 probe. These results provide a first indication that Abf2p influences differential organization of mtDNA sequences.

Journal Article.  6147 words.  Illustrated.

Subjects: Chemistry ; Biochemistry ; Bioinformatics and Computational Biology ; Genetics and Genomics ; Molecular and Cell Biology

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