NSF regulates membrane traffic along multiple pathways in Paramecium

Kissmehl, Roland; Froissard, Marine; Plattner, Helmut; Momayezi, Massoud; Cohen, Jean

NSF regulates membrane traffic along multiple pathways in Paramecium

Dateien

NSF_regulates_membrane_traffic_along_multiple_pathways_in_Paramecium.pdfGröße: 805.2 KBDownloads: 268

Datum

2002

Autor:innen

Kissmehl, Roland

Froissard, Marine

Plattner, Helmut

Momayezi, Massoud

Cohen, Jean

Projekt

Molekulare Spezifität von Membran-Interaktionen bei Paramecium

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Journal of Cell Science. 2002, 115(20), pp. 3935-3946. ISSN 0021-9533. eISSN 1477-9137. Available under: doi: 10.1242/jcs.00079

Zusammenfassung

N-ethylmaleimide (NEM)-sensitive factor (NSF), a regulator of soluble NSF attachment protein receptors (SNAREs), is required for vesicular transport in many eukaryotic cells. In the ciliated protozoon Paramecium, complex but well-defined transport routes exist, constitutive and regulated exocytosis, endocytosis, phagocytosis and a fluid excretory pathway through contractile vacuoles, that can all be studied independently at the whole cell level. To unravel the role of NSF and of the SNARE machinery in this complex traffic, we looked for NSF genes in Paramecium, starting from a partial sequence found in a pilot random sequencing project. We found two very similar genes, PtNSF1 and PtNSF2, which both seem to be expressed. Peptide-specific antibodies (Abs) recognize PtNSF as a 84 kDa band. PtNSF gene silencing results in decreasing phagocytotic activity, while stimulated exocytosis of dense core-vesicles (trichocysts), once firmly attached at the cell membrane, persists. Ultrastructural analysis of silenced cells shows deformation or disappearance of structures involved in membrane traffic. Aggregates of numerous small, smooth vesicles intermingled with branches of ER occur in the cytoplasm and are most intensely labeled with anti-NSF Ab-gold. Furthermore, elongated vesicles of ~30 nm diameter can be seen attached at cortical calcium storage compartments, the alveolar sacs, whose unknown biogenesis may thus be revealed. Involvement of PtNSF in some low frequency fusion events was visualized in non-silenced cells by immuno-fluorescence, after cautious permeabilization in the presence of ATP-γ-S and NEM. Our data document that PtNSF is involved in distinct pathways of vesicle traffic in Paramecium and that actual sensitivity to silencing is widely different, apparently dependent on the turnover of membrane-to-membrane attachment formation.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Schlagwörter

Ciliates, Paramecium, Secretion, Phagocytosis, Golgi, Endoplasmic reticulum

Zitieren

ISO 690

KISSMEHL, Roland, Marine FROISSARD, Helmut PLATTNER, Massoud MOMAYEZI, Jean COHEN, 2002. NSF regulates membrane traffic along multiple pathways in Paramecium. In: Journal of Cell Science. 2002, 115(20), pp. 3935-3946. ISSN 0021-9533. eISSN 1477-9137. Available under: doi: 10.1242/jcs.00079

BibTex

@article{Kissmehl2002regul-7804,
  year={2002},
  doi={10.1242/jcs.00079},
  title={NSF regulates membrane traffic along multiple pathways in Paramecium},
  number={20},
  volume={115},
  issn={0021-9533},
  journal={Journal of Cell Science},
  pages={3935--3946},
  author={Kissmehl, Roland and Froissard, Marine and Plattner, Helmut and Momayezi, Massoud and Cohen, Jean}
}

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