Tracking individual secretory vesicles during exocytosis reveals an ordered and regulated process.

Post-Golgi secretory vesicle trafficking is a coordinated process, with transport and regulatory mechanisms to ensure appropriate exocytosis. While the contributions of many individual regulatory proteins to this process are well studied, the timing and dependencies of events have not been defined. Here we track individual secretory vesicles and associated proteins in vivo during tethering and fusion in budding yeast.

Head-to-tail regulation is critical for the in vivo function of myosin V.

Cell organization requires regulated cargo transport along cytoskeletal elements. Myosin V motors are among the most conserved organelle motors and have been well characterized in both yeast and mammalian systems. Biochemical data for mammalian myosin V suggest that a head-to-tail autoinhibitory interaction is a primary means of regulation, but the in vivo significance of this interaction has not been studied.

Myosin-V is activated by binding secretory cargo and released in coordination with Rab/exocyst function.

Cell organization requires motor-dependent transport of specific cargos along cytoskeletal elements. How the delivery cycle is coordinated with other events is poorly understood. Here we define the in vivo delivery cycle of myosin-V in its essential function of secretory vesicle transport along actin cables in yeast.

A role for phospholipase A2 activity in membrane tubule formation and TGN trafficking.

We have investigated the role of phospholipase A(2) (PLA(2) ) enzymes in generating membrane tubules at the trans-Golgi network (TGN). Constitutive TGN membrane tubules and those induced by over-expressing kinase dead protein kinase D were inhibited by the PLA(2) inhibitors ONO-RS-082 (ONO) and bromoenol lactone. These antagonists also inhibited secretory delivery of both soluble and transmembrane cargoes.