Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei

Mari Valkonen (Corresponding author), Merja Penttilä, Markku Saloheimo, Nick Read

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

Abstract

Available data suggest that the basic principles of secretion obtained from studies on S. cerevisiae and animal cells also apply to filamentous fungi. Although considerable research has been performed on the secretion process in filamentous fungi, little is known about their secretory machinery. The transport of proteins between the cellular compartments occurs in membrane-bounded vesicles that bud from the donor and fuse with the acceptor compartment. Membrane bound proteins called SNAREs (Soluble N-ethylaleimide-sensitive factor Attachment protein REceptors) play a key role in membrane fusion. SNAREs have been found to be involved in most membrane fusion events in the cell. This study describes the cloning and characterization of a putative exocytotic t-SNARE (sso1) from Trichoderma reesei. We cloned the T. reesei homologue of Neurospora crassa nsyn2. This protein was labelled with the Cerulean fluorescent protein mCer and the fusion protein expressed in T. reesei. Transformants expressing the fusion protein exhibit a clear labelling of the plasma membrane. The fusion protein is uniformly distributed along the hyphae with no tip-focused gradient. A v-SNARE SNCI was labelled with mRFP to analyze its interaction with the SSOI protein using fluorescence resonance energy
transfer (FRET) imaging.
Original languageEnglish
Title of host publication8th European Conference on Fungal Genetics
Subtitle of host publicationBook of Abstracts
EditorsIrina S. Druzhinina, Alexey G. Kopchinskiy, Christian P. Kubicek
PublisherVienna University of Technology
ChapterVIII Protein Targeting and Secretion
Pages299
Publication statusPublished - 2006
Event8th European Conference on Fungal Genetics - Vienna, Austria
Duration: 8 Apr 200611 Apr 2006

Conference

Conference8th European Conference on Fungal Genetics
CountryAustria
CityVienna
Period8/04/0611/04/06

Fingerprint

Trichoderma reesei
protein secretion
image analysis
fungi
proteins
secretion
Neurospora crassa
transport proteins
hyphae
plasma membrane
buds
fluorescence
cells
receptors
animals

Cite this

Valkonen, M., Penttilä, M., Saloheimo, M., & Read, N. (2006). Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei. In I. S. Druzhinina, A. G. Kopchinskiy, & C. P. Kubicek (Eds.), 8th European Conference on Fungal Genetics: Book of Abstracts (pp. 299). [VIIIp-6] Vienna University of Technology.
Valkonen, Mari ; Penttilä, Merja ; Saloheimo, Markku ; Read, Nick. / Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei. 8th European Conference on Fungal Genetics: Book of Abstracts. editor / Irina S. Druzhinina ; Alexey G. Kopchinskiy ; Christian P. Kubicek. Vienna University of Technology, 2006. pp. 299
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abstract = "Available data suggest that the basic principles of secretion obtained from studies on S. cerevisiae and animal cells also apply to filamentous fungi. Although considerable research has been performed on the secretion process in filamentous fungi, little is known about their secretory machinery. The transport of proteins between the cellular compartments occurs in membrane-bounded vesicles that bud from the donor and fuse with the acceptor compartment. Membrane bound proteins called SNAREs (Soluble N-ethylaleimide-sensitive factor Attachment protein REceptors) play a key role in membrane fusion. SNAREs have been found to be involved in most membrane fusion events in the cell. This study describes the cloning and characterization of a putative exocytotic t-SNARE (sso1) from Trichoderma reesei. We cloned the T. reesei homologue of Neurospora crassa nsyn2. This protein was labelled with the Cerulean fluorescent protein mCer and the fusion protein expressed in T. reesei. Transformants expressing the fusion protein exhibit a clear labelling of the plasma membrane. The fusion protein is uniformly distributed along the hyphae with no tip-focused gradient. A v-SNARE SNCI was labelled with mRFP to analyze its interaction with the SSOI protein using fluorescence resonance energytransfer (FRET) imaging.",
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Valkonen, M, Penttilä, M, Saloheimo, M & Read, N 2006, Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei. in IS Druzhinina, AG Kopchinskiy & CP Kubicek (eds), 8th European Conference on Fungal Genetics: Book of Abstracts., VIIIp-6, Vienna University of Technology, pp. 299, 8th European Conference on Fungal Genetics, Vienna, Austria, 8/04/06.

Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei. / Valkonen, Mari (Corresponding author); Penttilä, Merja; Saloheimo, Markku; Read, Nick.

8th European Conference on Fungal Genetics: Book of Abstracts. ed. / Irina S. Druzhinina; Alexey G. Kopchinskiy; Christian P. Kubicek. Vienna University of Technology, 2006. p. 299 VIIIp-6.

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

TY - CHAP

T1 - Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei

AU - Valkonen, Mari

AU - Penttilä, Merja

AU - Saloheimo, Markku

AU - Read, Nick

N1 - CA2: TK402

PY - 2006

Y1 - 2006

N2 - Available data suggest that the basic principles of secretion obtained from studies on S. cerevisiae and animal cells also apply to filamentous fungi. Although considerable research has been performed on the secretion process in filamentous fungi, little is known about their secretory machinery. The transport of proteins between the cellular compartments occurs in membrane-bounded vesicles that bud from the donor and fuse with the acceptor compartment. Membrane bound proteins called SNAREs (Soluble N-ethylaleimide-sensitive factor Attachment protein REceptors) play a key role in membrane fusion. SNAREs have been found to be involved in most membrane fusion events in the cell. This study describes the cloning and characterization of a putative exocytotic t-SNARE (sso1) from Trichoderma reesei. We cloned the T. reesei homologue of Neurospora crassa nsyn2. This protein was labelled with the Cerulean fluorescent protein mCer and the fusion protein expressed in T. reesei. Transformants expressing the fusion protein exhibit a clear labelling of the plasma membrane. The fusion protein is uniformly distributed along the hyphae with no tip-focused gradient. A v-SNARE SNCI was labelled with mRFP to analyze its interaction with the SSOI protein using fluorescence resonance energytransfer (FRET) imaging.

AB - Available data suggest that the basic principles of secretion obtained from studies on S. cerevisiae and animal cells also apply to filamentous fungi. Although considerable research has been performed on the secretion process in filamentous fungi, little is known about their secretory machinery. The transport of proteins between the cellular compartments occurs in membrane-bounded vesicles that bud from the donor and fuse with the acceptor compartment. Membrane bound proteins called SNAREs (Soluble N-ethylaleimide-sensitive factor Attachment protein REceptors) play a key role in membrane fusion. SNAREs have been found to be involved in most membrane fusion events in the cell. This study describes the cloning and characterization of a putative exocytotic t-SNARE (sso1) from Trichoderma reesei. We cloned the T. reesei homologue of Neurospora crassa nsyn2. This protein was labelled with the Cerulean fluorescent protein mCer and the fusion protein expressed in T. reesei. Transformants expressing the fusion protein exhibit a clear labelling of the plasma membrane. The fusion protein is uniformly distributed along the hyphae with no tip-focused gradient. A v-SNARE SNCI was labelled with mRFP to analyze its interaction with the SSOI protein using fluorescence resonance energytransfer (FRET) imaging.

M3 - Conference abstract in proceedings

SP - 299

BT - 8th European Conference on Fungal Genetics

A2 - Druzhinina, Irina S.

A2 - Kopchinskiy, Alexey G.

A2 - Kubicek, Christian P.

PB - Vienna University of Technology

ER -

Valkonen M, Penttilä M, Saloheimo M, Read N. Imaging the last stages of protein secretion in the filamentous fungus Trichoderma reesei. In Druzhinina IS, Kopchinskiy AG, Kubicek CP, editors, 8th European Conference on Fungal Genetics: Book of Abstracts. Vienna University of Technology. 2006. p. 299. VIIIp-6