Domains of Gln3p interacting with karyopherins, Ure2p, and the target of rapamycin protein.
Gln3p is a GATA-type transcription factor responsive to the quality of nitrogen and carbon. In preferred nitrogen such as glutamine, Gln3p is phosphorylated and sequestered in the cytoplasm in a manner that is dependent on the target of rapamycin (TOR) protein and Ure2p. In nonpreferred nitrogen or nitrogen starvation, Gln3p is dephosphorylated and imported into the nucleus via karyopherin alpha/Srp1p. Upon reintroduction of preferred nitrogen, Gln3p is exported from the nucleus by Crm1p/Xpo1p. Although recent work has provided insights into Gln3p, a more detailed understanding is needed to elucidate the mechanism of its localization and function. In this study, we show that Gln3p contains canonical nuclear localization signal and nuclear export signal sequences necessary for its localization and interaction with its relevant karyopherins. In addition, we identify an N-terminal domain of Gln3p interacting with Ure2p and a C-terminal region for binding to TOR. Finally, we find a lysine/arginine-rich domain essential for the rapamycin-sensitive function, but dispensable for its localization. Our results reveal key domains of Gln3p important for its function and regulation.
Carvalho
John
J
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Zheng
X F Steven
XF
D001665:Binding Sites; D005979:Glutathione Peroxidase; D028884:Karyopherins / Q000378:metabolism*; D011328:Prions / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins / Q000032:analysis* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000378:metabolism*; D020123:Sirolimus / Q000378:metabolism*; D014157:Transcription Factors / Q000032:analysis* / Q000378:metabolism*
C071664:GLN3 protein, S cerevisiae; D028884:Karyopherins; D011328:Prions; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae; D020123:Sirolimus