Characterisation of two 14-3-3 genes from Trichoderma reesei: interactions with yeast secretory pathway components.
The 14-3-3 proteins are highly conserved, ubiquitously expressed proteins taking part in numerous cellular processes. Two genes encoding 14-3-3 proteins, ftt1 and ftt2, were isolated and characterised from the filamentous fungus Trichoderma reesei. FTTI showed the highest sequence identity (98% at the amino acid level) to the Trichoderma harzianum protein Th1433. FTTII is relatively distinct from FTTI, showing approximately 75% identity to other fungal 14-3-3 proteins. Despite their sequence divergence, both of the T. reesei ftt genes were equally able to complement the yeast bmh1 bmh2 double disruption. The T. reesei ftt genes were also found to be quite closely linked in the genomic DNA. A C-terminally truncated version of ftt1 (ftt1DeltaC) was first isolated as a multicopy suppressor of the growth defect of the temperature-sensitive yeast secretory mutant sec15-1. Overexpression of ftt1DeltaC also suppressed the growth defect of sec2-41, sec3-101, and sec7-1 strains. Overexpression of ftt1DeltaC in sec2-41 and sec15-1 strains could also rescue the secretion of invertase at the restrictive temperatures, and overexpression of full-length ftt1 enhanced invertase secretion by wild-type yeast cells. These findings strongly suggest that the T. reesei ftt1 has a role in protein secretion.
Vasara
Tuija
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VTT Biotechnology, P.O. Box 1500, Tietotie 2, FIN-02044 VTT, Espoo, Finland.
Keränen
Sirkka
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Penttilä
Merja
M
Saloheimo
Markku
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D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006026:Glycoside Hydrolases / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D017386:Sequence Homology, Amino Acid; D014242:Trichoderma / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics*; D043324:beta-Fructofuranosidase
D048948:14-3-3 Proteins; D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; D014446:Tyrosine 3-Monooxygenase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase