Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

Abstract

The machinery for trafficking proteins through the secretory pathway is well conserved in eukaryotes, but remains poorly characterized in filamentous fungi. Secretion is believed to be highly polarized in fungal hyphae and to mainly occur from hyphal tips. We describe the isolation of the snc1 and sso1 genes encoding exocytic SNARE proteins from Trichoderma reesei. The encoded SNCI protein can complement Snc protein depletion in S. cerevisiae whilst the T. reesei SSOI protein was unable to complement depletion of its yeast homologues. The localization and interactions of the T. reesei SNARE proteins were studied with advanced fluorescence imaging methods using fluorescent fusions of the SNARE proteins. The SSOI and SNCI proteins co-localized in sterol-independent clusters on the plasma membrane in sub-apical but not apical hyphal regions. The v-SNARE SNCI (but not the t-SNARE SSOI) localized to the apical vesicle cluster within the Spitzenkörper of the growing hyphal tips when expressed from the homologous T. reesei cbh1 promoter. Using fluorescence lifetime imaging microscopy (FLIM) and fluorescence energy transfer (FRET) analysis, we quantified the interactions between these proteins with high spatial resolution in living cells. Our data showed that the site of SNARE complex formation between these proteins is on the plasma membrane of non-growing hyphae in old sub-peripheral regions of the colony, but that there is no interaction between the proteins in growing hyphal tips in at the colony margin. These findings suggest spatially distinct sites of exocytosis within filamentous fungi and the existence of multiple exocytic SNAREs which are functionally and spatially segregated
Original languageEnglish
Title of host publication3rd European Federation of Biotechnology Conference
Subtitle of host publicationPhysiology of Yeasts and Filamentous Fungi PYFF3
Place of PublicationEspoo
PublisherVTT Technical Research Centre of Finland
Pages132
ISBN (Electronic)978-951-38-6314-2
ISBN (Print)978-951-38-6313-5
Publication statusPublished - 2007
Event3rd European Federation of Biotechnology Conference : Physiology of Yeasts and Filamentous Fungi - Helsinki, Finland
Duration: 13 Jun 200716 Jun 2007

Publication series

NameVTT Symposium
PublisherVTT
Number245
ISSN (Print)0357-9387
ISSN (Electronic)1455-0873

Conference

Conference3rd European Federation of Biotechnology Conference
Abbreviated titlePYFF3
CountryFinland
CityHelsinki
Period13/06/0716/06/07

Fingerprint

Trichoderma reesei
fungi
proteins
fluorescence
hyphae
complement
plasma membrane
image analysis
protein depletion
protein transport
exocytosis
energy transfer
sterols
eukaryotic cells
microscopy
promoter regions
SNARE proteins
secretion
yeasts
genes

Cite this

Valkonen, M., Saloheimo, M., Penttilä, M., Read, N. D., & Duncan, R. R. (2007). Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei. In 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 (pp. 132). [P76] Espoo: VTT Technical Research Centre of Finland. VTT Symposium, No. 245
Valkonen, Mari ; Saloheimo, Markku ; Penttilä, Merja ; Read, N.D. ; Duncan, R.R. / Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei. 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 . Espoo : VTT Technical Research Centre of Finland, 2007. pp. 132 (VTT Symposium; No. 245).
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abstract = "The machinery for trafficking proteins through the secretory pathway is well conserved in eukaryotes, but remains poorly characterized in filamentous fungi. Secretion is believed to be highly polarized in fungal hyphae and to mainly occur from hyphal tips. We describe the isolation of the snc1 and sso1 genes encoding exocytic SNARE proteins from Trichoderma reesei. The encoded SNCI protein can complement Snc protein depletion in S. cerevisiae whilst the T. reesei SSOI protein was unable to complement depletion of its yeast homologues. The localization and interactions of the T. reesei SNARE proteins were studied with advanced fluorescence imaging methods using fluorescent fusions of the SNARE proteins. The SSOI and SNCI proteins co-localized in sterol-independent clusters on the plasma membrane in sub-apical but not apical hyphal regions. The v-SNARE SNCI (but not the t-SNARE SSOI) localized to the apical vesicle cluster within the Spitzenk{\"o}rper of the growing hyphal tips when expressed from the homologous T. reesei cbh1 promoter. Using fluorescence lifetime imaging microscopy (FLIM) and fluorescence energy transfer (FRET) analysis, we quantified the interactions between these proteins with high spatial resolution in living cells. Our data showed that the site of SNARE complex formation between these proteins is on the plasma membrane of non-growing hyphae in old sub-peripheral regions of the colony, but that there is no interaction between the proteins in growing hyphal tips in at the colony margin. These findings suggest spatially distinct sites of exocytosis within filamentous fungi and the existence of multiple exocytic SNAREs which are functionally and spatially segregated",
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Valkonen, M, Saloheimo, M, Penttilä, M, Read, ND & Duncan, RR 2007, Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei. in 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 ., P76, VTT Technical Research Centre of Finland, Espoo, VTT Symposium, no. 245, pp. 132, 3rd European Federation of Biotechnology Conference , Helsinki, Finland, 13/06/07.

Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei. / Valkonen, Mari; Saloheimo, Markku; Penttilä, Merja; Read, N.D.; Duncan, R.R.

3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 . Espoo : VTT Technical Research Centre of Finland, 2007. p. 132 P76 (VTT Symposium; No. 245).

Research output: Chapter in Book/Report/Conference proceedingConference abstract in proceedingsScientific

TY - CHAP

T1 - Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei

AU - Valkonen, Mari

AU - Saloheimo, Markku

AU - Penttilä, Merja

AU - Read, N.D.

AU - Duncan, R.R.

PY - 2007

Y1 - 2007

N2 - The machinery for trafficking proteins through the secretory pathway is well conserved in eukaryotes, but remains poorly characterized in filamentous fungi. Secretion is believed to be highly polarized in fungal hyphae and to mainly occur from hyphal tips. We describe the isolation of the snc1 and sso1 genes encoding exocytic SNARE proteins from Trichoderma reesei. The encoded SNCI protein can complement Snc protein depletion in S. cerevisiae whilst the T. reesei SSOI protein was unable to complement depletion of its yeast homologues. The localization and interactions of the T. reesei SNARE proteins were studied with advanced fluorescence imaging methods using fluorescent fusions of the SNARE proteins. The SSOI and SNCI proteins co-localized in sterol-independent clusters on the plasma membrane in sub-apical but not apical hyphal regions. The v-SNARE SNCI (but not the t-SNARE SSOI) localized to the apical vesicle cluster within the Spitzenkörper of the growing hyphal tips when expressed from the homologous T. reesei cbh1 promoter. Using fluorescence lifetime imaging microscopy (FLIM) and fluorescence energy transfer (FRET) analysis, we quantified the interactions between these proteins with high spatial resolution in living cells. Our data showed that the site of SNARE complex formation between these proteins is on the plasma membrane of non-growing hyphae in old sub-peripheral regions of the colony, but that there is no interaction between the proteins in growing hyphal tips in at the colony margin. These findings suggest spatially distinct sites of exocytosis within filamentous fungi and the existence of multiple exocytic SNAREs which are functionally and spatially segregated

AB - The machinery for trafficking proteins through the secretory pathway is well conserved in eukaryotes, but remains poorly characterized in filamentous fungi. Secretion is believed to be highly polarized in fungal hyphae and to mainly occur from hyphal tips. We describe the isolation of the snc1 and sso1 genes encoding exocytic SNARE proteins from Trichoderma reesei. The encoded SNCI protein can complement Snc protein depletion in S. cerevisiae whilst the T. reesei SSOI protein was unable to complement depletion of its yeast homologues. The localization and interactions of the T. reesei SNARE proteins were studied with advanced fluorescence imaging methods using fluorescent fusions of the SNARE proteins. The SSOI and SNCI proteins co-localized in sterol-independent clusters on the plasma membrane in sub-apical but not apical hyphal regions. The v-SNARE SNCI (but not the t-SNARE SSOI) localized to the apical vesicle cluster within the Spitzenkörper of the growing hyphal tips when expressed from the homologous T. reesei cbh1 promoter. Using fluorescence lifetime imaging microscopy (FLIM) and fluorescence energy transfer (FRET) analysis, we quantified the interactions between these proteins with high spatial resolution in living cells. Our data showed that the site of SNARE complex formation between these proteins is on the plasma membrane of non-growing hyphae in old sub-peripheral regions of the colony, but that there is no interaction between the proteins in growing hyphal tips in at the colony margin. These findings suggest spatially distinct sites of exocytosis within filamentous fungi and the existence of multiple exocytic SNAREs which are functionally and spatially segregated

M3 - Conference abstract in proceedings

SN - 978-951-38-6313-5

T3 - VTT Symposium

SP - 132

BT - 3rd European Federation of Biotechnology Conference

PB - VTT Technical Research Centre of Finland

CY - Espoo

ER -

Valkonen M, Saloheimo M, Penttilä M, Read ND, Duncan RR. Spatially segregated snare protein interactions in filamentous fungus Trichoderma reesei. In 3rd European Federation of Biotechnology Conference: Physiology of Yeasts and Filamentous Fungi PYFF3 . Espoo: VTT Technical Research Centre of Finland. 2007. p. 132. P76. (VTT Symposium; No. 245).