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Nucleic Acids Research, 2001, Vol. 29, No. 22 4736-4743
© 2001 Oxford University Press

Sea urchin mtDBP is a two-faced transcription termination factor with a biased polarity depending on the RNA polymerase

Patricio Fernandez-Silva, Paola Loguercio Polosa1, Marina Roberti1, Barbara Di Ponzio1, Maria Nicola Gadaleta1, Julio Montoya and Palmiro Cantatore1,*

Departamento de Bioquimica y Biologia Molecular y Celular, Universidad de Zaragoza, Miguel Servet 177, E-50013 Zaragoza, Spain and 1Dipartimento di Biochimica e Biologia Molecolare, Università di Bari, and the Centro Studi sui Mitocondri e Metabolismo Energetico, CNR, Bari, Via Orabona 4, 70125 Bari, Italy

The sea urchin mitochondrial displacement (D)-loop binding protein mtDBP has been previously identified and cloned. The polypeptide (348 amino acids) displays a significant homology with the human mitochondrial transcription termination factor mTERF. This similarity, and the observation that the 3' ends of mitochondrial RNAs coded by opposite strands mapped in correspondence of mtDBP-binding sites, suggested that mtDBP could function as transcription termination factor in sea urchin mitochondria. To investigate such a role we tested the capability of mtDBP bound to its target sequence in the main non-coding region to affect RNA elongation by mitochondrial and bacteriophage T3 and T7 RNA polymerases. We show that mtDBP was able to terminate transcription bidirectionally when initiated by human mitochondrial RNA polymerase but only unidirectionally when initiated by T3 or T7 RNA polymerases. Time-course experiments indicated that mtDBP promotes true transcription termination rather than transcription pausing. These results indicate that mtDBP is able to function as a bipolar transcription termination factor in sea urchin mitochondria. The functional significance of such an activity could be linked to the previously proposed dual role of the protein in modulating mitochondrial DNA transcription and replication.

* To whom correspondence should be addressed. Tel: +39 080 5443378; Fax: +39 080 5443403; Email: p.cantatore{at}biologia.uniba.it The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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