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Free Content Protein trafficking to the cell wall occurs through mechanisms distinguishable from default sorting in tobacco

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Summary

The secretory pathway in plants involves sustained traffic to the cell wall, as matrix components, polysaccharides and proteins reach the cell wall through the endomembrane system. We studied the secretion pattern of cell-wall proteins in tobacco protoplasts and leaf epidermal cells using fluorescent forms of a pectin methylesterase inhibitor protein (PMEI1) and a polygalacturonase inhibitor protein (PGIP2). The two most representative protein fusions, secGFP–PMEI1 and PGIP2–GFP, reached the cell wall by passing through ER and Golgi stacks but using distinct mechanisms. secGFP–PMEI1 was linked to a glycosylphosphatidylinositol (GPI) anchor and stably accumulated in the cell wall, regulating the activity of the endogenous pectin methylesterases (PMEs) that are constitutively present in this compartment. A mannosamine-induced non-GPI-anchored form of PMEI1 as well as a form (PMEI1–GFP) that was unable to bind membranes failed to reach the cell wall, and accumulated in the Golgi stacks. In contrast, PGIP2–GFP moved as a soluble cargo protein along the secretory pathway, but was not stably retained in the cell wall, due to internalization to an endosomal compartment and eventually the vacuole. Stable localization of PGIP2 in the wall was observed only in the presence of a specific fungal endopolygalacturonase ligand in the cell wall. Both secGFP–PMEI1 and PGIP2–GFP sorting were distinguishable from that of a secreted GFP, suggesting that rigorous and more complex controls than the simple mechanism of bulk flow are the basis of cell-wall growth and differentiation.
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Keywords: GPI anchor; PGIP2; PMEI1; cell-wall trafficking; endocytosis; secretion pathway

Document Type: Research Article

Affiliations: 1: Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, 73100 Lecce, Italy 2: Dipartimento di Biologia e Biotecnologie ‘C. Darwin’, Università‘La Sapienza’, 00185 Roma, Italy

Publication date: January 1, 2011

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