ADP ribosylation factor regulates spectrin binding to the Golgi complex

Homologues of two major components of the well-characterized erythrocyte plasma-membrane-skeleton, spectrin (a not-yet-cloned isoform, beta I Sigma* spectrin) and ankyrin (Ank(G119) and an approximate to 195-kDa ankyrin), associate with the Golgi complex, ADP ribosylation factor (ARF) is a small G protein that controls the architecture and dynamics of the Golgi by mechanisms that remain incompletely understood. We find that activated ARF stimulates the in vitro association of beta I Sigma* spectrin with a Golgi fraction, that the Golgi-associated beta I Sigma* spectrin contains epitopes characteristic of the beta I Sigma 2 spectrin pleckstrin homology (PPI) domain known to bind phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)), and that ARF recruits beta I Sigma* spectrin by inducing increased PtdInsP(2) levels in the Golgi, The stimulation of spectrin binding by ARF is independent of its ability to stimulate phospholipase D or to recruit coat proteins (COP)-I and can be blocked by agents that sequester PtdInsP(2), We postulate that a PH domain within beta I Sigma*: Golgi spectrin binds PtdInsP(2) and acts as a regulated docking site for spectrin on the Golgi, Agents that block the binding of spectrin to the Golgi, either by blocking the PH domain interaction or a constitutive Golgi binding site within spectrin's membrane association domain I, inhibit the transport of vesicular stomatitis virus G protein from endoplasmic reticulum to the medial compartment of the Golgi complex, Collectively, these results suggest that the Golgi-spectrin skeleton plays a central role in regulating the structure and function of this organelle.