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(Received for publication, July 11, 1996, and in revised form, September 9, 1996)
From the Departments of Terminal segments (telomeres) of linear
mitochondrial DNA (mtDNA) molecules of the yeast Candida
parapsilosis consist of large sequence units repeated in tandem.
The extreme ends of mtDNA terminate with a 5
Volume 272, Number 5,
Issue of January 31, 1997
pp. 3049-3056
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
ka
,
Genetics and
¶ Biochemistry, Faculty of Natural Sciences, Comenius
University, 842 15 Bratislava, Slovakia, and the 
Institut
Curie, Section de Recherche, Batiment 110, Centre Universitaire
Paris XI, 91 405 Orsay, France
single-stranded overhang
of about 110 nucleotides. We identified and purified a
i
ochondrial
elomere-
inding 
rotein (mtTBP)
that specifically recognizes a synthetic oligonucleotide derived from
the extreme end of this linear mtDNA. MtTBP is highly resistant to
protease and heat treatments, and it protects the telomeric probe from
degradation by various DNA-modifying enzymes. Resistance of the complex
to bacterial alkaline phosphatase suggests that mtTBP binds the very
end of the molecule. We purified mtTBP to near homogeneity using DNA
affinity chromatography based on the telomeric oligonucleotide
covalently bound to Sepharose. Sodium dodecyl sulfate-polyacrylamide
gel electrophoretic analysis of the purified fractions revealed the
presence of a protein with an apparent molecular mass of ~15 kDa. UV
cross-linking and gel filtration chromatography experiments suggested
that native mtTBP is probably a homo-oligomer. MtTBP of C. parapsilosis is the first identified protein that specifically
binds to telomeres of linear mitochondrial DNA.
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