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Abstract
To ensure their homeostasis and sustain differentiated functions, cells continuously transport diverse cargos to various cell compartments and in particular to the cell surface. Secreted proteins are transported along intracellular routes from the endoplasmic reticulum through the Golgi complex before reaching the plasma membrane along microtubule tracks. Using a synchronized secretion assay, we report here that exocytosis does not occur randomly at the cell surface but on localized hotspots juxtaposed to focal adhesions. Although microtubules are involved, the RAB6-dependent machinery plays an essential role. We observed that, irrespective of the transported cargos, most post-Golgi carriers are positive for RAB6 and that its inactivation leads to a broad reduction of protein secretion. RAB6 may thus be a general regulator of post-Golgi secretion.
Highlights
To reach the cell surface, secreted proteins are transported along interacting/CAST family member 1; known as RAB6IP2).intracellular routes from the endoplasmic reticulum (ER) ELKS is an effector of the Golgi-associated Ras-related protein 6 through the Golgi complex
Using the RUSH assay, we studied the synchronous secretion of diverse cargos: collagen type X (ColX), VSV-G, secretory soluble EGFP, gp135, the glycosylphosphatidylinositol-anchored proteins (GPI-APs) cluster of differentiation 59 (CD59) and placenta alkaline phosphatase (PLAP), and TNFα
About 35 min after biotin addition, most of ColX had been secreted into the medium, and almost no signal remained in cells
Summary
To reach the cell surface, secreted proteins are transported along interacting/CAST family member 1; known as RAB6IP2).intracellular routes from the endoplasmic reticulum (ER) ELKS is an effector of the Golgi-associated Ras-related protein 6 through the Golgi complex. Cargos exit the Golgi complex in (RAB6) GTPase (Monier et al, 2002), which regulates several transport carriers that use microtubules to be addressed rapidly anterograde and retrograde trafficking pathways to and from to the plasma membrane before exocytosis. Transmembrane the Golgi complex, as well as Golgi homeostasis (Goud, 1999; proteins are exposed at the plasma membrane while solu- White et al, 1999; Mallard et al, 2002; Grigoriev et al, 2007). Whether de- particular, RAB6 was shown to be involved in the targeting of livery of cargos occurs randomly or at specific sites of the plasma post-Golgi vesicles containing the secretory markers vesicular membrane is still unclear, and the mechanisms that direct ex- stomatitis virus glycoprotein Microtubules were described to be protein), and neuropeptide Y (NPY; a soluble protein) to ELKS-
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