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Volume 271, Number 23, Issue of June 7, 1996 pp. 13537-13541
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Isolation of a 25-kDa Protein Binding to a Curved DNA Upstream the Origin of the L Strand Replication in the Rat Mitochondrial Genome

(Received for publication, January 7, 1996, and in revised form, February 28, 1996)

Gemma Gadaleta , Domenica D'Elia , Lara Capaccio , Cecilia Saccone and Gabriella Pepe

From the Centro di Studio sui Mitocondri e Metabolismo Energetico-CNR and Department of Biochemistry and Molecular Biology, University of Bari, via Orabona 4A, 70126 Bari, Italy

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.


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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.
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