Confirmation of the subcellular interactions of the kinase complexes AnSte11-Ste7 and AnSte7-Fus3 by BIFC system.

Abstract

<p>(A) Interaction of N-EYFP::AnSte11 [SteC] and C-EYFP::AnSte7 [MkkB] proteins in the hyphal cells. AnSte11-Ste7 kinase complexes are located at the plasma membrane, septal connections, the hyphal tip and partially nuclear envelope. The upper panel shows the localization of the YFP signal in comparison to nuclear mRFP::Histone2A fluorescence. Lower panel displays the YFP signal emitting cells stained with membrane dye FM4-64. (B) Physical interaction of N-EYFP::AnSte7 [MkkB] with C-EYFP::AnFus3 [MpkB] proteins in the fungal cells. (C) Quantification of the subcellular locations of the AnSte11-Ste7 complexes that are often present at the hyphal tip, plasma membrane, septum and nuclear envelope. N:50 fungal cells were counted in triplicate. Standard deviations are presented as vertical bars. (D) Subcellular locations of the AnSte7-Fus3 interactions. AnSte7-Fus3 complexes hardly localize to the septum and are found more on the nuclear envelope. (E) Assembly of the AnSte11-AnSte7 and AnSte7-AnFus3 complexes on the surface of vesicles of asexual conidiophores. Arrows indicate the growth directions of the metulae initials on the vesicles. V; vesicle, S; stalk. Size of the scale bars is 10 µm.</p

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