Isolation of a 25-kDa protein binding to a curved DNA upstream the origin of the L strand replication in the rat mitochondrial genome.
The presence of a curved DNA sequence in the gene for the NADH-dehydrogenase subunit 2 of rat mitochondrial genome, upstream from the origin of the light strand replication have been demonstrated through theoretical analysis and experimental approaches. Gel retardation assays showed that this structure makes a complex with a protein component extracted from the mitochondrial matrix. The isolation and purification of this protein is reported. With a Sepharose CL-6B and magnetic DNA affinity chromatography a polypeptide was purified to homogeneity having 25-kDa mass as shown by gel electrophoresis. To functionally characterize this protein, its capability to bind to other sequences of the homologous or heterologous DNA and to specific riboprobes was also investigated. A role for this protein as a trans-acting agent required for the expression of the mammalian mitochondrial genome is suggested.
Gadaleta
G
G
Centro di Studio sui Mitocondri e Metabolismo Energetico-CNR, University of Bari, via Orabona 4A, 70126 Bari, Italy.
D'Elia
D
D
Capaccio
L
L
Saccone
C
C
Pepe
G
G
D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D004261:DNA Replication; D004272:DNA, Mitochondrial / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D016678:Genome; D066298:In Vitro Techniques; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009245:NADH Dehydrogenase / Q000737:chemistry / Q000235:genetics; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D051381:Rats; D018741:Replication Origin
D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D009245:NADH Dehydrogenase