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Factors beyond enolase 2 and mitochondrial lysyl-tRNA synthetase precursor are required for tRNA import into yeast mitochondria

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Abstract

In yeast, the import of tRNALys with CUU anticodon (tRK1) relies on a complex mechanism where interaction with enolase 2 (Eno2p) dictates a deep conformational change of the tRNA. This event is believed to mask the tRNA from the cytosolic translational machinery to re-direct it towards the mitochondria. Once near the mitochondrial outer membrane, the precursor of the mitochondrial lysyl-tRNA synthetase (preMsk1p) takes over enolase to carry the tRNA within the mitochondrial matrix, where it is supposed to participate in translation following correct refolding. Biochemical data presented in this report focus on the role of enolase. They show that despite the inability of Eno2p alone to form a complex with tRK1, mitochondrial import can be recapitulated in vitro using fractions of yeast extracts sharing either recombinant or endogenous yeast Eno2p as one of the main components. Taken together, our data suggest the existence of a protein complex containing Eno2p that is involved in RNA mitochondrial import.

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Abbreviations

EMSA:

electrophoretic mobility shift assay

Eno2p:

enolase 2

ITC:

isothermal titration microcalorimetry

preMsk1p:

precursor of mitochondrial lysyl-tRNA synthetase

tRK1:

tRNALys with CUU anticodon

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Author information

Authors and Affiliations

  1. GMGM, CNRS − University of Strasbourg, UMR 7156, 67081, Strasbourg, France

    M. V. Baleva, N. Entelis, I. Tarassov & B. Masquida

  2. Lomonosov Moscow State University, Faculty of Biology, 119991, Moscow, Russia

    M. V. Baleva & P. Kamenski

  3. IGBMC, CNRS − University of Strasbourg, UMR 7104, 67404, Illkirch, France

    M. Meyer

  4. Immanuil Kant Baltic Federal University, Institute of Living Systems, 236038, Kaliningrad, Russia

    P. Kamenski

Authors
  1. M. V. Baleva
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  2. M. Meyer
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  3. N. Entelis
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  4. I. Tarassov
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  5. P. Kamenski
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  6. B. Masquida
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Corresponding author

Correspondence to B. Masquida.

Additional information

Published in Russian in Biokhimiya, 2017, Vol. 82, No. 11, pp. 1681–1695.

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Baleva, M.V., Meyer, M., Entelis, N. et al. Factors beyond enolase 2 and mitochondrial lysyl-tRNA synthetase precursor are required for tRNA import into yeast mitochondria. Biochemistry Moscow 82, 1324–1335 (2017). https://doi.org/10.1134/S0006297917110104

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  • DOI: https://doi.org/10.1134/S0006297917110104

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