Biochemical and functional characterization of ORF138, a mitochondrial protein responsible for Ogura cytoplasmic male sterility in Brassiceae.
In cytoplasmic male sterility (CMS), original mitochondrial genes contribute to sex determinism by provoking pollen abortion. The function of the encoded proteins remains unclear. We studied the ORF138 protein, responsible for the 'Ogura' CMS, which is both used in hybrid seed production and present in natural populations. We analyzed the biochemical and structural properties of this protein in male-sterile plants and in E. coli. We showed that this protein spontaneously forms dimers in vitro. Truncated variants of the protein, containing either the hydrophobic or the hydrophilic moiety, also spontaneously dimerize. By fractionating mitochondria, we showed that ORF138 was strongly associated with the inner mitochondrial membrane of male-sterile plants. Our results also strongly suggest that ORF138 forms oligomers in male-sterile plant mitochondria. In E. coli, ORF138 was associated with the plasma membrane, as shown by membrane fractionation, and formed oligomers. The production of this protein strongly inhibited bacterial growth, but not by inhibiting respiration. The observed toxic effects required both the hydrophilic and hydrophobic moieties of the protein.
Duroc
Yann
Y
Station de Génétique et d'Amélioration des Plantes, Institut Jean-Pierre Bourgin, INRA, Route de Saint-Cyr, 78026 Versailles cedex, France.
Gaillard
Catherine
C
Hiard
Sophie
S
Defrance
Marie-Christine
MC
Pelletier
Georges
G
Budar
Françoise
F
D001483:Base Sequence; D001937:Brassica / Q000378:metabolism*; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051479:Plant Infertility; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012098:Reproduction; D015183:Restriction Mapping
D017931:DNA Primers; D024101:Mitochondrial Proteins; D010940:Plant Proteins