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. 2012 Dec 28;7(12):e52956. doi: 10.1371/journal.pone.0052956

Figure 6. Localization of Mdy2 mutants with defective nuclear localization, Mdy2-ΔNLS, and nuclear export, Mdy2-ΔNES.

Figure 6

(A) Mdy2 localizes mainly to the nucleus at 28°C. Mdy2 localize to cytoplasmic granules following heat stress. Mdy2 co-localize with Pab1 following mild and robust heat stress. GFP-Mdy2 and Pab1-RFP was visualized by fluorescence microscopy in a mdy2Δ strain transformed with plasmids containing GFP-Mdy2 and Pab1-RFP (upper panel) or GFP-Mdy2 and Dcp2-RFP (lower panel) after a temperature shift to 38°C (middle panel) and 46°C (right panel). (B) Localization of Mdy2-ΔNLS and Mdy2-ΔNES mutant proteins at permissive temperature. Exponentially growing mdy2Δ cells carrying wild type Mdy2 (MDY2), empty vector (mdy), NLS and NES deletion constructs (mdy2-ΔNLS and mdy2-ΔNES, respectively) fused to the C-terminus of GFP protein in CEN plasmids and expressed under the control of MDY2-own promoter. GFP-fusion proteins were localized by direct fluorescence and nuclear DNA was stained by DAPI. (C) Localization of Mdy2-ΔNLS and Mdy2-ΔNES mutant proteins during heat shock-induced stress. Localization pattern for Mdy2-ΔNLS and Mdy2-ΔNES mutant proteins as indicated in (A) during heat shock-induced stress. Localization of GFP-Mdy2 and GFP-Mdy2-NES to the cytoplasmic granules following heat stress is shown in upper and lower panel.