Fig. 4.

PMA1 localization and function. (A) Immuno-staining of fixed yeast cells (using anti-HA antibodies) in a strain expressing Pma1 fused to the HA peptide. This ‘native’ Pma1 localizes exclusively to the plasma membrane, with none evident in the vacuole. Reproduced with permission from Mason et al. (2014). (B) Live cell imaging of yeast expressing a Pma1-EGFP fusion protein, expressed from the endogenous Pma1 promoter. A significant amount of fluorescence is observed in the vacuole in addition to that present at the plasma membrane. (C) Comparison of the growth of yeast expressing untagged Pma1 (1), Pma1 C-terminally tagged with mCherry (2), or Pma1 C-terminally tagged with SpyTag (3). Strains are streaked on media containing 2% galactose, so the Pma1-ST expressing strain is also expressing SpyoIPD-EGFP. For both tagged strains, Pma1 is expressed under control of its native promoter, and the tagged copy of the strain is the only copy of the protein present. The strain expressing Pma1-mCherry fusion exhibits a significant growth defect, whereas the other two strains grow equally well.