Spatially segregated SNARE protein interactions in living fungal cells

Mari Valkonen (Corresponding Author), Eric R. Kalkman, Markku Saloheimo, Merja Penttilä, Nick D. Read, Rory R. Duncan

Research output: Contribution to journalArticleScientificpeer-review

38 Citations (Scopus)

Abstract

The machinery for trafficking proteins through the secretory pathway is well conserved in eukaryotes, from fungi to mammals. We describe the isolation of the snc1, sso1, and sso2 genes encoding exocytic SNARE proteins from the filamentous fungus Trichoderma reesei. The localization and interactions of the T. reesei SNARE proteins were studied with advanced fluorescence imaging methods. The SSOI and SNCI proteins co-localized in sterol-independent clusters on the plasma membrane in subapical but not apical hyphal regions. The vesicle SNARE SNCI also localized to the apical vesicle cluster within the Spitzenkörper of the growing hyphal tips. Using fluorescence lifetime imaging microscopy and Foerster resonance energy transfer analysis, we quantified the interactions between these proteins with high spatial resolution in living cells. Our data showed that the site of ternary SNARE complex formation between SNCI and SSOI or SSOII, respectively, is spatially segregated. SNARE complex formation could be detected between SNCI and SSOI in subapical hyphal compartments along the plasma membrane, but surprisingly, not in growing hyphal tips, previously thought to be the main site of exocytosis. In contrast, SNCI·SSOII complexes were found exclusively in growing apical hyphal compartments. These findings demonstrate spatially distinct sites of plasma membrane SNARE complex formation in fungi and the existence of multiple exocytic SNAREs, which are functionally and spatially segregated. This is the first demonstration of spatially regulated SNARE interactions within the same membrane.

Original languageEnglish
Pages (from-to)22775-22785
JournalJournal of Biological Chemistry
Volume282
Issue number31
DOIs
Publication statusPublished - 3 Aug 2007
MoE publication typeA1 Journal article-refereed

Fingerprint

SNARE Proteins
Cell membranes
Fungi
Optical Imaging
Cell Membrane
Fluorescence
Imaging techniques
Proteins
Trichoderma
Gene encoding
Mammals
Secretory Pathway
Energy Transfer
Exocytosis
Sterols
Protein Transport
Eukaryota
Energy transfer
Machinery
Microscopy

Keywords

  • metabolic pathways
  • gene expression
  • genes
  • SNARE
  • SNARE proteins
  • Trichoderma reesei
  • filamentous fungi

Cite this

Valkonen, Mari ; Kalkman, Eric R. ; Saloheimo, Markku ; Penttilä, Merja ; Read, Nick D. ; Duncan, Rory R. / Spatially segregated SNARE protein interactions in living fungal cells. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 31. pp. 22775-22785.
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Spatially segregated SNARE protein interactions in living fungal cells. / Valkonen, Mari (Corresponding Author); Kalkman, Eric R.; Saloheimo, Markku; Penttilä, Merja; Read, Nick D.; Duncan, Rory R.

In: Journal of Biological Chemistry, Vol. 282, No. 31, 03.08.2007, p. 22775-22785.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Spatially segregated SNARE protein interactions in living fungal cells

AU - Valkonen, Mari

AU - Kalkman, Eric R.

AU - Saloheimo, Markku

AU - Penttilä, Merja

AU - Read, Nick D.

AU - Duncan, Rory R.

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KW - gene expression

KW - genes

KW - SNARE

KW - SNARE proteins

KW - Trichoderma reesei

KW - filamentous fungi

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SP - 22775

EP - 22785

JO - Journal of Biological Chemistry

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