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Truncated presequences of mitochondrial F1-ATPase β subunit from Nicotiana plumbaginifolia transport CAT and GUS proteins into mitochondria of transgenic tobacco

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Abstract

The mitochondrial F1-ATPase β subunit (ATPase-β) of Nicotiana plumbaginifolia is nucleus-encoded as a precursor containing an NH2-terminal extension. By sequencing the mature N. tabacum ATPase-β, we determined the length of the presequence, viz. 54 residues. To define the essential regions of this presequence, we produced a series of 3′ deletions in the sequence coding for the 90 NH2-terminal residues of ATPase-β. The truncated sequences were fused with the chloramphenicol acetyl transferase (cat) and β-glucuronidase (gus) genes and introduced into tobacco plants. From the observed distribution of CAT and GUS activity in the plant cells, we conclude that the first 23 amino-acid residues of ATPase-β remain capable of specifically targeting reporter proteins into mitochondria. Immunodetection in transgenic plants and in vitro import experiments with various CAT fusion proteins show that the precursors are processed at the expected cleavage site but also at a cryptic site located in the linker region between the presequence and the first methionine of native CAT.

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Authors and Affiliations

  1. Unité de Biochimie Physiologique, University of Louvain, Place Croix du Sud, 2-20, B-1348, Louvain-la-Neuve, Belgium

    François Chaumont, Marcio de Castro Silva Filho, Didier Thomas, Serge Leterme & Marc Boutry

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  1. François Chaumont
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  2. Marcio de Castro Silva Filho
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  3. Didier Thomas
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  4. Serge Leterme
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  5. Marc Boutry
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Chaumont, F., de Castro Silva Filho, M., Thomas, D. et al. Truncated presequences of mitochondrial F1-ATPase β subunit from Nicotiana plumbaginifolia transport CAT and GUS proteins into mitochondria of transgenic tobacco. Plant Mol Biol 24, 631–641 (1994). https://doi.org/10.1007/BF00023559

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