Class 1 Reversibly Glycosylated Polypeptides Are Plasmodesmal-Associated Proteins Delivered to Plasmodesmata via the Golgi Apparatus
| Autor: | Guy Sagi, Dana Guenoune-Gelbart, Bernard L. Epel, Aviva Katz |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Yellow fluorescent protein
Glycosylation Recombinant Fusion Proteins Molecular Sequence Data Arabidopsis Golgi Apparatus Cell Communication Plant Science Plasmodesma Biology Green fluorescent protein symbols.namesake chemistry.chemical_compound Cell Wall Tobacco Research Articles Conserved Sequence Glycoproteins Brefeldin A Sequence Homology Amino Acid Arabidopsis Proteins Callose Plasmodesmata Membrane Transport Proteins Cell Biology Golgi apparatus Molecular biology Fusion protein Blot Protein Transport chemistry symbols biology.protein Carrier Proteins Peptides |
| Zdroj: | The Plant Cell. 17:1788-1800 |
| ISSN: | 1532-298X |
| DOI: | 10.1105/tpc.105.031823 |
| Popis: | SE-WAP41, a salt-extractable 41-kD wall-associated protein that is associated with walls of etiolated maize (Zea mays) seedlings and is recognized by an antiserum previously reported to label plasmodesmata and the Golgi, was cloned, sequenced, and found to be a class 1 reversibly glycosylated polypeptide (C1RGP). Protein gel blot analysis of cell fractions with an antiserum against recombinant SE-WAP41 showed it to be enriched in the wall fraction. RNA gel blot analysis along the mesocotyl developmental axis and during deetiolation demonstrates that high SE-WAP41 transcript levels correlate spatially and temporally with primary and secondary plasmodesmata (Pd) formation. All four of the Arabidopsis thaliana C1RGP proteins, when fused to green fluorescent protein (GFP) and transiently expressed in tobacco (Nicotiana tabacum) epidermal cells, display fluorescence patterns indicating they are Golgi- and plasmodesmal-associated proteins. Localization to the Golgi apparatus was verified by colocalization of transiently expressed AtRGP2 fused to cyan fluorescence protein together with a known Golgi marker, Golgi Nucleotide Sugar Transporter 1 fused to yellow fluorescent protein (GONST1:YFP). In transgenic tobacco, AtRGP2:GFP fluorescence is punctate, is present only in contact walls between cells, and colocalizes with aniline blue–stained callose present around Pd. In plasmolyzed cells, AtRGP2:GFP remains wall embedded, whereas GONST1:YFP cannot be found embedded in cell walls. This result implies that the targeting to Pd is not due to a default pathway for Golgi-localized fusion proteins but is specific to C1RGPs. Treatment with the Golgi disrupting drug Brefeldin A inhibits Pd labeling by AtRGP2:GFP. Integrating these data, we conclude that C1RGPs are plasmodesmal-associated proteins delivered to plasmodesmata via the Golgi apparatus. |
| Databáze: | OpenAIRE |
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