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. 2008 Aug 1;7(10):1674–1684. doi: 10.1128/EC.00151-08

FIG. 1.

FIG. 1.

Internal loop 6 and the C-terminal region of Dtr1p are essential for PSM localization. (A) Cartoon of predicted topology of Dtr1p in the lipid bilayer and the truncation/deletion mutant proteins used in this study. Predicted amino acid positions of transmembrane domains are as follows: TM1, 111 to 133; TM2, 148 to 160; TM3, 177 to 200; TM4, 203 to 226; TM5, 237 to 260; TM6, 265 to 287; TM7, 367 to 382; TM8, 404 to 426; TM9, 447 to 469; TM10, 473 to 495; TM11, 508 to 531; and TM12, 540 to 562 (http://db.yeastgenome.org/cgi-bin/protein). (B) Localization of full-length Dtr1p-GFP, Dtr1NΔp-GFP, and Dtr1C15Δp-GFP in sporulating cells (Y7730, Y7747, and Y7733, respectively) expressing Mpc54p-RFP, a MOP marker. Dtr1C15Δp-GFP failed to be transported to the PSM and showed cytoplasmic puncta in the vicinity of Mpc54p-RFP signals. (C) Dtr1Loop6Δp-RFP localized to the ER, as demonstrated by colocalization with the ER marker Sec63p-GFP, in sporulating cells (Y7657). Sec63p-GFP showed dispersed staining in the cytoplasm in addition to ER staining. (D) FM4-64 staining of sporulating cells expressing Dtr1C15Δp-GFP (Y7637) 5 h after the induction of sporulation. FM4-64 stained PM (0 min), was internalized into the cytoplasm (10 min), and was transported to vacuoles (30 min). Dtr1C15Δp-GFP puncta did not overlap with FM4-64 signals at any time point. (E) Localization of Dtr1C15Δp-GFP in sporulating cells expressing mRFP-Sed5p (Y8044) or Sec7p-dsRed (Y7744). Dtr1C15Δp-GFP did not colocalize with Sec7p-dsRed, but signals overlapped with mRFP-Sed5p. Bars, 5 μm.