The presequence of a precursor to the δ-subunit of sweet potato mitochondrial F1ATPase is not sufficient for the transport of β-glucuronidase (GUS) into mitochondria of tobacco, rice and yeast cells

Tetsuya Kimura, Shin Takeda, Junko Kyozuka, Tadashi Asahi, Ko Shimamoto, Kenzo Nakamura

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

A precursor to the δ-subunit of sweet potato mitochondrial F1ATPase (pre-F1δ) has an amino-terminal (N-terminal) presequence of 45 amino acid residues and its N-terminal 18 residues may form an amphiphilic a-helix, which is typical of mitochondrial targeting signals [Kimura et al. (1990) J. Biol. Chem. 265: 6079]. Fusion genes consisting of sequences that encoded the 25 (DG25), 46 (DG46) and 73 (DG73) N-terminal amino acids from pre-F1δ fused to the N-terminus of the coding sequence of bacterial β-glucuronidase (GUS) were placed downstream of the 35S promoter of cauliflower mosaic virus and used to transform suspension-cultured tobacco cells, rice calli and tobacco plants. Fusion genes were also placed downstream of the yeast GAL10 promoter and introduced into Saccharomyces cerevisiae cells. In these transformed cells, only the DG73 GUS-fusion protein was transported into mitochondria and subjected to proteolytic cleavage of the presequence. Neither transport to mitochondria nor processing of the presequence of the DG46 GUS-fusion protein, which contained the entire presequence and the processing site, occurred in either plant or yeast cells. These results indicate that the presequence of pre-F1δ is not sufficient for the transport of the GUS protein into mitochondria in tobacco, rice and yeast cells. The requirement for the longer polypeptide from pre-F1δ in the transport of the GUS protein into mitochondria could be due either to the lack of sufficient information for mitochondrial targeting within the presequence or to the nature of the passenger protein, GUS, used in this study.

Original languageEnglish
Pages (from-to)345-355
Number of pages11
JournalPlant and Cell Physiology
Volume34
Issue number2
Publication statusPublished - 1993 Mar 1
Externally publishedYes

Keywords

  • Genetic transformation
  • Mitochondria
  • Protein transport
  • β-Glucuronidase (GUS)

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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