Rvs161p and Rvs167p, the two yeast amphiphysin homologs, function together in vivo.
Mutations in RVS161 and RVS167, the two yeast amphiphysin homologs, cause very similar growth phenotypes, a depolarized actin cytoskeleton, and a defect in the internalization step of endocytosis. Rvs161p and Rvs167p have been shown to interact in the two-hybrid system, but their localization in the cell may be different thus raising the question whether the interaction is physiologically relevant. Here we demonstrate that the two proteins function together in vivo. We find that the steady state level of Rvs167p is strongly reduced in an rvs161Delta strain. Similarly, the level of Rvs161p is strongly reduced in an rvs167Delta strain. We demonstrate that these reduced protein levels at steady state are due to a decreased stability of either Rvs protein in the absence of the other protein. Furthermore, we find that the amount and ratio of Rvs161p and Rvs167p are critical parameters for receptor-mediated endocytosis. In addition, by using the two-hybrid system we show that the interaction of Rvs167p with actin is not abolished in an abp1Delta strain suggesting that Abp1p is not essential for this interaction.
Lombardi
R
R
Biozentrum of the University of Basel, CH-4056 Basel, Switzerland.
Riezman
H
H
D000199:Actins / Q000378:metabolism; D002455:Cell Division; D003598:Cytoskeletal Proteins; D004705:Endocytosis; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D008840:Microfilament Proteins; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D020798:Two-Hybrid System Techniques; D015003:Yeasts
C062612:ABP1 protein, S cerevisiae; D000199:Actins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; C089181:RVS161 protein, S cerevisiae; C082282:RVS167 protein, S cerevisiae; D011956:Receptors, Cell Surface; D029701:Saccharomyces cerevisiae Proteins; C075637:amphiphysin