A Rab1 GTPase Is Required for Transport between the Endoplasmic Reticulum and Golgi Apparatus and for Normal Golgi Movement in Plants

TLDR

The authors developed a GFP‑based assay to study membrane trafficking along the secretory pathway in plants. They expressed a dominant‑negative AtRab1b(N121I) mutant to block ER‑to‑Golgi transport, observing GFP marker accumulation, and showed rescue by co‑expressing wild‑type AtRab1b. The results demonstrate that AtRab1b is essential for ER‑to‑Golgi transport and that its loss impairs Golgi movement, effects reversed by wild‑type AtRab1b.

Abstract

We describe a green fluorescent protein (GFP)–based assay for investigating membrane traffic on the secretory pathway in plants. Expression of AtRab1b(N121I), predicted to be a dominant inhibitory mutant of the Arabidopsis Rab GTPase AtRab1b, resulted in accumulation of a secreted GFP marker in an intracellular reticulate compartment reminiscent of the endoplasmic reticulum. This accumulation was alleviated by coexpressing wild-type AtRab1b but not AtRab8c. When a Golgi-targeted and N-glycosylated variant of GFP was coexpressed with AtRab1b(N121I), the variant also accumulated in a reticulate network and an endoglycosidase H–sensitive population appeared. Unexpectedly, expression of AtRab1b(N121I), but not of the wild-type AtRab1b, resulted in a reduction or cessation of vectorial Golgi movement, an effect that was reversed by coexpression of the wild type. We conclude that AtRab1b function is required for transport from the endoplasmic reticulum to the Golgi apparatus and suggest that this process may be coupled to the control of Golgi movement.