A putative mitochondrial fission gene from the ectomycorrhizal ascomycete Tuber borchii Vittad.: cloning, characterisation and phylogeny.
Mitochondrial binary division is a complex process occurring in multiple steps, mediated by several proteins. In Saccharomyces cerevisiae, a mitochondrial membrane protein, Fis1p, is required for the proper assembly of the mitochondrial division apparatus. In this study, we report the cloning, characterisation and phylogenetic analysis of Tbfis1, a gene from the ectomycorrhizal ascomycetous truffle Tuber borchii, encoding for an orthologue of S. cerevisiae Fis1p. The Tbfis1 coding region consists of a 468-nucleotide open reading frame interrupted by four introns, which encodes for a polypeptide of 155 amino acids, having a predicted transmembrane domain structure typical of the Fis1p Family. Southern blot analysis revealed that Tbfis1 is a single-copy gene in the T. borchii genome. Tbfis1 is highly expressed during the first stages of T. borchii fruit body ripening, while its expression decreases during T. borchii mycelium ageing. Also, Virtual Northern blot analysis revealed Tbfis1 expression in the symbiotic phase of the fungus life cycle. Phylogenetic analysis allowed the identification of Tbfis1 orthologues in filamentous fungi, yeasts, plants, worms, flies and mammals, indicating that the function of the protein coded by this gene has been conserved during evolution.
Guidi
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C
Istituto di Chimica Biologica Giorgio Fornaini, Università degli Studi di Urbino Carlo Bo, via Saffi 2, PU 61029 Urbino, Italy.
Zeppa
S
S
Barbieri
E
E
Zambonelli
A
A
Polidori
E
E
Potenza
L
L
Stocchi
V
V
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000235:genetics*; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D017931:DNA Primers; D040461:Gene Components; D015870:Gene Expression; D007438:Introns / Q000235:genetics; D016013:Likelihood Functions; D008565:Membrane Proteins / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics* / Q000502:physiology; D024101:Mitochondrial Proteins / Q000235:genetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010802:Phylogeny; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D017931:DNA Primers; C464859:FIS1 protein, S cerevisiae; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins