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Neuhaus, Jean-Marc

Nom
Neuhaus, Jean-Marc
Affiliation principale
Fonction
Professeur.e émérite
Email
jean-marc.neuhaus@unine.ch
Identifiants
https://libra.unine.ch/handle/123456789/300
_
0000-0003-3288-101X

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  • Publication
    Métadonnées seulement
    Two glycosylated vacuolar GFPs are new markers for ER-to-vacuole sorting
    (2013)
    Stigliano, Egidio
    ;
    Faraco, Marianna
    ;
    Neuhaus, Jean-Marc 
    ;
    Montefusco, Anna
    ;
    Dalessandro, Giuseppe
    ;
    Piro, Gabriella
    ;
    Di Sansebastiano, Gian-Pietro
    Vacuolar Sorting Determinants (VSDs) have been extensively studied in plants but the mechanisms for the accumulation of storage proteins in somatic tissues are not yet fully understood. In this work we used two mutated versions of well-documented vacuolar fluorescent reporters, a GFP fusion in frame with the C-terminal VSD of tobacco chitinase (GFPChi) and an N-terminal fusion in frame with the sequence-specific VSD of the barley cysteine protease aleurain (AleuGFP). The GFP sequence was mutated to present an N-glycosylation site at the amino-acid position 133. The reporters were transiently expressed in Nicotiana tabacum protoplasts and agroinfiltrated in Nicotiana benthamiana leaves and their distribution was identical to that of the non-glycosylated versions. With the glycosylated GFPs we could highlight a differential ENDO-H sensitivity and therefore differential glycan modifications. This finding suggests two different and independent routes to the vacuole for the two reporters. BFA also had a differential effect on the two markers and further, inhibition of COPII trafficking by a specific dominant-negative mutant (NtSar1h74l) confirmed that GFPChi transport from the ER to the vacuole is not fully dependent on the Golgi apparatus.
  • Publication
    Accès libre
    Vacuolar system distribution in Arabidopsis tissues, visualized using GFP fusion proteins
    (2003-06-01)
    Flückiger, Ricardo
    ;
    De Caroli, Monica
    ;
    Piro, Gabriella
    ;
    Dalessandro, Giuseppe
    ;
    Neuhaus, Jean-Marc 
    ;
    Di Sansebastiano, Gian-Pietro
    Green fluorescent protein (GFP) allows the direct visualization of gene expression and the subcellular localization of fusion proteins in living cells. The localization of different GFP fusion proteins in the secretory system was studied in stably transformed Arabidopsis plants cv. Wassilewskaja. Secreted GFP (SGFP) and GFP retained in the ER (GFP-KDEL) confirmed patterns already known, but two vacuolar GFPs (GFP-Chi and Aleu-GFP) labelled the Arabidopsis vacuolar system for the first time, the organization of which appears to depend on cell differentiation. GFP stability in the vacuoles may depend on pH or degradation, but these vacuolar markers can, nevertheless, be used as a tool for physiological studies making these plants suitable for mutagenesis and gene-tagging experiments.