title: Glutamic acid decarboxylase gene disruption reveals signalling pathway(s) governing complex morphogenic and metabolic events in Trichoderma atroviride.
abstract: Glutamate decarboxylase (GAD) catalyses decarboxylation of glutamate to gamma-aminobutyrate (GABA) in a metabolic pathway connected to citrate cycle and known as GABA shunt. The gene (gad) was disrupted in Trichoderma atroviride CCM F-534 and viable mutants were characterized. Two of them were found to arise by homologous recombination and were devoid of both GAD activity and GABA. Mutants grew slower as compared to the wild type (F534). In the submerged culture, mutants developed less CO2 and consumed less O2 than the F534 without changing their respiratory quotients. Hyphae of mutants were more ramified than those of F534. Their ramification, in contrast to F534, was not increased by cyclosporin A, a drug causing hyphae ramification of several fungi and which is a calcineurin/cyclophilin inhibitor, or by FK506. Rapamycin, which is a cyclophilin but not calcineurin inhibitor, had a different effect on hyphae ramification in F534 and mutants. To examine the presence of GABA receptors in the fungus the effect of mammalian GABA-receptor modulators, such as bicuculline, gabapentin or carbamazepine on fungal morphology were investigated. Conidia of mutants germinated in a multipolar manner more frequently (up to 80 %) than those of F534. This trait was modified with cyclosporine A, FK506 and GABA receptor modulators in a different manner. Transport of chlorides, an intimate feature of GABA-regulated receptors/channels in animal cells, was measured in vegetative mycelia by means (36)Cl(-) uptake. It was significantly reduced in gad mutants. The results suggest that T. atroviride possesses a signalling pathway that involves GABA, putative GABA receptor(s), calcineurin, target of rapamycin and chloride transporter(s) to regulate physiological functions.
authors.affiliation: Department of Biochemistry and Microbiology, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37, Bratislava, Slovakia, lubosniznansky@gmail.com.
authors.forename: Luboš
authors.initials: L
authors.lastname: Nižňanský
authors.forename: Svetlana
authors.initials: S
authors.lastname: Kryštofová
authors.forename: Peter
authors.initials: P
authors.lastname: Vargovič
authors.forename: Michal
authors.initials: M
authors.lastname: Kaliňák
authors.forename: Martin
authors.initials: M
authors.lastname: Simkovič
authors.forename: Ludovít
authors.initials: L
authors.lastname: Varečka
mesh_terms: D000935:Antifungal Agents / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D016572:Cyclosporine / Q000378:metabolism; D055786:Gene Knockout Techniques; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism*; D025301:Hyphae / Q000254:growth & development; D053858:Metabolic Networks and Pathways; D010100:Oxygen / Q000378:metabolism; D015398:Signal Transduction; D020123:Sirolimus / Q000378:metabolism; D014242:Trichoderma / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D002245:Carbon Dioxide; D016572:Cyclosporine; D005968:Glutamate Decarboxylase; D010100:Oxygen; D020123:Sirolimus