title: A voltage-gated pore for translocation of tRNA.
abstract: Very little is known about how nucleic acids are translocated across membranes. The multi-subunit RNA Import Complex (RIC) from mitochondria of the kinetoplastid protozoon Leishmania tropica induces translocation of tRNAs across artificial or natural membranes, but the nature of the translocation pore remains unknown. We show that subunits RIC6 and RIC9 assemble on the membrane in presence of subunit RIC4A to form complex R3. Atomic Force Microscopy of R3 revealed particles with an asymmetric surface groove of ∼20 nm rim diameter and ∼1 nm depth. R3 induced translocation of tRNA into liposomes when the pH of the medium was lowered to ∼6 in the absence of ATP. R3-mediated tRNA translocation could also be induced at neutral pH by a K(+) diffusion potential with an optimum of 60-70 mV. Point mutations in the Cys2-His2 Fe-binding motif of RIC6, which is homologous to the respiratory Complex III Fe-S protein, abrogated import induced by low pH but not by K(+) diffusion potential. These results indicate that the R3 complex forms a pore that is gated by a proton-generated membrane potential and that the Fe-S binding region of RIC6 has a role in proton translocation. The tRNA import complex of L. tropica thus contains a novel macromolecular channel distinct from the mitochondrial protein import pore that is apparently involved in tRNA import in some species.
authors: Genetic Engineering Laboratory, Molecular and Human Genetics Division, CSIR-Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Calcutta 700 032, India.
authors: Sandip
authors: S
authors: Koley
authors: Samit
authors: S
authors: Adhya
mesh_terms: D020816:Amino Acid Motifs; D004058:Diffusion; D006863:Hydrogen-Ion Concentration; D007895:Leishmania tropica / Q000235:genetics* / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D018625:Microscopy, Atomic Force; D008928:Mitochondria / Q000378:metabolism; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010316:Particle Size; D011188:Potassium / Q000378:metabolism; D011522:Protons; D034443:RNA Transport; D012343:RNA, Transfer / Q000235:genetics* / Q000378:metabolism
chemical_list: D011522:Protons; D012343:RNA, Transfer; D011188:Potassium
keywords: AFM; Membrane potential; Mutagenesis; Translocation; tRNA