Mislocalization of large ARF-GEFs as a potential mechanism for BFA resistance in COG-deficient cells.

Defects in subunits of the conserved oligomeric Golgi (COG) complex represent a growing subset of congenital disorders of glycosylation (CDGs). In addition to altered protein glycosylation and vesicular trafficking, Cog-deficient patient fibroblasts exhibit a striking delay in the Golgi-disrupting effects of brefeldin A (BFA). Despite the diagnostic value of this BFA resistance, the molecular basis of this response is not known. To investigate potential mechanisms of resistance, we analyzed the localization of the large ARF-GEF, GBF1, in several Cog-deficient cell lines. Our results revealed mislocalization of GBF1 to non-Golgi compartments, in particular the ERGIC, within these cells. Biochemical analysis of GBF1 in control and BFA-treated fibroblasts demonstrated that the steady-state level and membrane recruitment is not substantially affected by COG deficiency, supporting a role for the COG complex in the localization but not membrane association of GBF1. We also showed that pretreatment of fibroblasts with bafilomycin resulted in a GBF1-independent BFA resistance that appears additive with the resistance associated with COG deficiency. These data provide new insight into the mechanism of BFA resistance in Cog-deficient cells by suggesting a role for impaired ARF-GEF localization.