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Molecular and Cellular Biology, August 2003, p. 5217-5224, Vol. 23, No. 15
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.15.5217-5224.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

"Ping-Pong" Interactions between Mitochondrial tRNA Import Receptors within a Multiprotein Complex

Subhendra Nath Bhattacharyya, Saibal Chatterjee, Srikanta Goswami, Gayatri Tripathi, Sailendra Nath Dey, and Samit Adhya*

Indian Institute of Chemical Biology, Calcutta 700032, India

Received 27 January 2003/ Returned for modification 4 April 2003/ Accepted 7 May 2003

The mitochondrial genomes of a wide variety of species contain an insufficient number of functional tRNA genes, and translation of mitochondrial mRNAs is sustained by import of nucleus-encoded tRNAs. In Leishmania, transfer of tRNAs across the inner membrane can be regulated by positive and negative interactions between them. To define the factors involved in such interactions, a large multisubunit complex (molecular mass, ~640 kDa) from the inner mitochondrial membrane of the kinetoplastid protozoon Leishmania, consisting of ~130-Å particles, was isolated. The complex, when incorporated into phospholipid vesicles, induced specific, ATP- and proton motive force-dependent transfer of Leishmania tRNATyr as well as of oligoribonucleotides containing the import signal YGGYAGAGC. Moreover, allosteric interactions between tRNATyr and tRNAIle were observed in the RNA import complex-reconstituted system, indicating the presence of primary and secondary tRNA binding sites within the complex. By a combination of antibody inhibition, photochemical cross-linking, and immunoprecipitation, it was shown that binding of tRNAIle to a 21-kDa component of the complex is dependent upon tRNATyr, while binding of tRNATyr to a 45-kDa component is inhibited by tRNAIle. This "ping-pong" mechanism may be an effective means to maintain a balanced tRNA pool for mitochondrial translation.

* Corresponding author. Mailing address: Indian Institute of Chemical Biology, 4 Raja S. C. Mullick Rd., Calcutta 700032, India. Phone: 91 33 2473 3491, ext. 136. Fax: 91 33 2473 5197. E-mail: sadhya{at}iicb.res.in.

Molecular and Cellular Biology, August 2003, p. 5217-5224, Vol. 23, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.15.5217-5224.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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