Ternary complex formation of Ino2p-Ino4p transcription factors and Apl2p adaptin beta subunit in yeast.
Yeast Ino2p-Ino4p heterodimeric complex is well known as a transcriptional activator for the genes regulated by inositol and choline, such as the INO1 gene. Apl2p is a large subunit of the yeast adaptin complex, an adaptor complex required for the clathrin coat to bind to the membrane. We found that Ino2p, Ino4p, and Apl2p form a ternary complex. This interaction was initially observed in a yeast two-hybrid study and subsequently verified by co-immunoprecipitation. Ino2p and Ino4p bind to Apl2p in the same region of Apl2p, viz., at the middle part and the C-terminal part. Ino2p and Ino4p bind to Apl2p independently, but more strongly when both are present. Furthermore, a disruption of APL2 together with INO2 or INO4 rendered yeast cells sensitive to oxidative stress. INO2-APL2 double disruptants also showed growth inability in non-fermentable carbon sources, such as glycerol. These results indicate a genetic interaction between APL2, INO2 and INO4 and uncovere novel functions of the Ino2p-Ino4p-Apl2p complex in yeast.
Nikawa
Jun-ichi
J
Department of Bioscience and Bioinformatics, Faculty of Computer Science and Systems Engineering, Kyushu Institute of Technology, Iizuka, Fukuoka, Japan. nikawa@bio.kyutech.ac.jp
Yata
Masako
M
Motomura
Miki
M
Miyoshi
Nobutaka
N
Ueda
Tsuyoshi
T
Hisada
Daisuke
D
D033966:Adaptor Protein Complex beta Subunits / Q000235:genetics / Q000378:metabolism*; D051792:Basic Helix-Loop-Helix Proteins; D002244:Carbon / Q000378:metabolism; D002966:Clathrin / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D006861:Hydrogen Peroxide / Q000494:pharmacology; D018384:Oxidative Stress; D010641:Phenotype; D011485:Protein Binding; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D033966:Adaptor Protein Complex beta Subunits; D051792:Basic Helix-Loop-Helix Proteins; D002244:Carbon; D002966:Clathrin; D006861:Hydrogen Peroxide; D021122:Protein Subunits; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C429285:APL2 protein, S cerevisiae; C095723:INO2 protein, S cerevisiae; C063311:INO4 protein, S cerevisiae