Intracellular localization of an active green fluorescent protein-tagged Pho84 phosphate permease in Saccharomyces cerevisiae

Green fluorescent protein (GFP) from Aerquorea victroia in was used as an i n vivo reporter protein when fused to the carboxy-terminus of the Pho84 pho sphate permease of Saccharomyces cerevisiae. Both components of the fusion protein displayed their native functions and revealed a cellular Localizati on and degradation of the Pho84-GFP chimera consistent with the behavior of the wild-type Pho84 protein. The GFP-tagged chimera allowed for a detectio n of conditions under which the Pho84 transporter is localized to its funct ional environment, i.e, the plasma membrane, and conditions linked to reloc ation of the protein to the vacuole for degradation. By use of the methodol ogy described, GFP should be useful in studies of localization and degradat ion also of other membrane proteins in vivo. (C) 1999 Federation of Europea n Biochemical Societies.