Fig 1.

C. neoformans Wsp1 is a conserved effector of Cin1 in actin organization. (A) In vivo localization of fluorescence-labeled fusion proteins. The GFP-CIN1 and DsRed-WSP1 fusion genes were expressed in the cin1 mutant strain. GFP-Cin1 is enriched in the plasma membrane as punctae, and DsRed-Wsp1 exhibits similar localization. DIC, differential interference contrast. (B) Endocytic uptake of FM4-64 in the wild-type (WT), cin1, and wsp1 strains. The cin1 and wsp1 mutants were defective in endocytosis, in agreement with previous studies (38, 39). (C) FM4-64 uptake by the cin1 mutant expressing either the wild-type WSP1 gene or the WSP1-B-GBD allele driven by the constitutively active GPD promoter. Wsp1-B-GBD expression results in internalization of FM4-64 as random patches and partial restoration of endocytosis in 60 min (indicated by arrows), in contrast to the pattern observed with the wild-type Wsp1 protein. Cells were prepared for fluorescence microscopy as described in the text.