Figure 5
Heterologous expression of N-terminal fragments of CHS5 from diverse fungal lineages: S. cerevisiae (ScChs5*), K. lactis (KlChs5*), C. albicans (CaChs5*), U. maydis (UmChs5*) and M. circinelloides (McChs5*). (A) Localization of N-terminal fragments of Chs5 from the indicated proteins tagged with GFP and co-localization of these fragments with the TGN marker Sec. 7-mRuby2. Images were acquired after induction in galactose media for the indicated times. (B) Co-localization of heterologously expressed versions of Chs5 with the Chs6 ChAP. The plasmids containing the indicated version of Chs5* were transformed in a S. cerevisiae chs5∆ mutant with a chromosomally tagged version of CHS6 (CHS6-mCherry). Proteins were expressed in galactose media for one hour as indicated. Note the apparent co-localization of green and red signals in some strains, indicative of an effective recruitment of Chs6-mCh from the cytoplasm by the heterologous expressed Chs5* fragments. (C) Complementation of the chs5∆ phenotypes by the heterologous expressed proteins. Note the alleviation of some phenotypes upon overexpression on galactose media. Overexpression of McChs5*-GFP showed deleterious effects on growth, therefore the complementation test is unreliable. All test were preformed in a S. cerevisiae chs5∆ strain grown overnight in raffinose media and later plated directly on galactose plates after appropriated dilution. See text and Table 1 for additional details on the interpretation of the Figure.
