ThPTR2, a di/tri-peptide transporter gene from Trichoderma harzianum.
The generation of a wide ESTs library and database from Trichoderma harzianum CECT 2413 was the base for identifying the gene ThPTR2, coding for a PTR family di/tri-peptide transporter. The deduced protein sequence of the ThPTR2 gene showed the conserved motifs and also the 12 transmembrane domains typical of the PTR transporters. The highest level of ThPTR2 expression was found when the fungus was grown in chitin as sole carbon source. We also found that ThPTR2 expression was increased when Trichoderma interacted directly in solid medium with the plant-pathogenic fungus Botrytis cinerea, showing that ThPTR2 is involved in the mycoparasitic process. Additionally, its expression was triggered by nitrogen starvation and a higher level of expression was also found when Trichoderma was grown in secondary nitrogen sources like allantoin, yeast extract, and urea. However, no difference was found when Trichoderma was grown in presence or absence of glucose as carbon source. Strain T34-15, a transformant that overexpressed the ThPTR2 gene, showed about a 2-fold increase in the uptake of the dipeptide Leu-Leu. Additionally, two transformants from the strain Trichoderma longibrachiatum T52 that overexpressed ThPTR2 were also studied, confirming the role of this gene in peptide transport. Other homologous genes to ThPTR2 were identified in other Trichoderma strains. ThPTR2 is the first experimentally confirmed PTR family transporter gene from filamentous fungi.
Vizcaíno
J A
JA
Spanish-Portuguese Center of Agricultural Research (CIALE), Departamento de Microbiología y Genética, Universidad de Salamanca, Edificio Departamental lab 208, Plaza Doctores de la Reina s/n, 37007 Salamanca, Spain.
Cardoza
R E
RE
Hauser
M
M
Hermosa
R
R
Rey
M
M
Llobell
A
A
Becker
J M
JM
Gutiérrez
S
S
Monte
E
E
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D020171:Botrytis / Q000254:growth & development; D002244:Carbon / Q000378:metabolism; D002686:Chitin / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004151:Dipeptides / Q000378:metabolism; D020224:Expressed Sequence Tags; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D005786:Gene Expression Regulation; D015966:Gene Expression Regulation, Fungal; D026901:Membrane Transport Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic; D014242:Trichoderma / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D004271:DNA, Fungal; D004151:Dipeptides; D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D002686:Chitin; C038361:leucylleucine; D002244:Carbon; C043912:peptide permease; D009584:Nitrogen