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Eukaryotic Cell, June 2003, p. 560-568, Vol. 2, No. 3
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.3.560-568.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

RBP38, a Novel RNA-Binding Protein from Trypanosomatid Mitochondria, Modulates RNA Stability

Sandro Sbicego,1,{dagger} Juan D. Alfonzo,2,{ddagger} Antonio M. Estévez,1,§ Mary Anne T. Rubio,2,{ddagger} Xuedong Kang,1 Christoph W. Turck,1,|| Marian Peris,1,# and Larry Simpson1,2*

Howard Hughes Medical Institute,1 Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, California 900952

Received 20 December 2002/ Accepted 29 January 2003

We describe here the isolation and characterization of a novel RNA-binding protein, RBP38, from Leishmania tarentolae mitochondria. This protein does not contain any known RNA-binding motifs and is highly conserved among the trypanosomatids, but no homologues were found in other organisms. Recombinant LtRBP38 binds single and double-stranded (ds) RNA substrates with dissociation constants in the 100 nM range, as determined by fluorescence polarization analysis. Downregulation of expression of the homologous gene, TbRBP38, in procyclic Trypanosoma brucei by using conditional dsRNA interference resulted in 80% reduction of steady-state levels of RNAs transcribed from both maxicircle and minicircle DNA. In organello pulse-chase labeling experiments were used to determine the stability of RNAs in mitochondria that were depleted of TbRBP38. The half-life of metabolically labeled RNA decreased from ~160 to ~60 min after depletion. In contrast, there was no change in transcriptional activity. These observations suggest a role of RBP38 in stabilizing mitochondrial RNA.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, 6780 MacDonald Research Laboratories, Los Angeles, CA 90095-1662. Phone: (310) 825-4215. Fax: (310) 206-8967. E-mail: simpson{at}kdna.ucla.edu.

{dagger} Present address: Ivoclar Vivadent AG, FL-9494 Schaan, Liechtenstein.

{ddagger} Present address: Department of Microbiology, Ohio State University, Columbus, OH 43210-12922.

§ Present address: Instituto de Parasitologia y Biomedicina "Lopez-Neyra," CSIC Ventanilla, 11, 18001 Granada, Spain.

|| Present address: Max Planck Institute of Psychiatry, Molecular, Cellular, Clinical Proteomics, D-80804 Munich, Germany.

# Present address: Molecular and Cell Biology Department and Howard Hughes Medical Institute, University of California, Berkeley, CA 94720.

Eukaryotic Cell, June 2003, p. 560-568, Vol. 2, No. 3
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.3.560-568.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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