Role of heterotrimeric G proteins in polarized membrane transport

MDCK cells maintain the polarized distribution of surface proteins mainly by sorting the newly synthesized proteins in the trans-Golgi network (TGN). In order to identify the components of the putative sorting machinery and to study factors that affect the sorting process, we have developed an in vitro system that reconstitutes the transport of viral glycoproteins from the TGN to the apical or basolateral surface. We have used this system to study effects of membrane impermeable reagents (such as peptides and antibodies) on the polarized transport. We observed that reagents affecting the stimulatory class (Gs) of heterotrimeric GTP binding proteins (G proteins) influenced the apical but not the basolateral transport. In contrast, reagents specific for the inhibitory class of G proteins (Gi) affected the basolateral but not the apical transport. These results show that the heterotrimeric G proteins differentially regulate the two pathways of polarized transport. The G proteins may regulate the process of polarized sorting of proteins in a fashion analogous to their role in signal transduction by providing a communication link with the cytosolic side of the membrane.