Cell-specific phosphorylation of Zfhep transcription factor.
Zinc finger homeodomain enhancer-binding protein (Zfhep/Zfhx1a) is a transcription factor essential for immune system development, skeletal patterning, and life. Regulation of the interleukin-2 gene in T cells has been suggested to depend on post-translational processing of Zfhep, however, no modifications of Zfhep are known. Here we demonstrate that Zfhep is present in both hyperphosphorylated and hypophosphorylated forms. Western blot analysis demonstrates two forms of Zfhep with different mobilities. Differences in phosphorylation are sufficient to explain the difference in mobilities. Zfhep is primarily phosphorylated on Ser and Thr residues since PP2A dephosphorylates the slower mobility band. Treatment of nuclear extract with O-GlcNAcase did not detect O-linked sugar. Importantly, post-translational processing is cell-specific. Doublets of Zfhep were detected in five cell lines, whereas 6 cell lines contain only, or predominantly, non-phosphorylated Zfhep, and Saos-2 cells contain predominantly the phosphorylated form. These data provide the first demonstration that Zfhep is post-translationally modified.
Costantino
Mary E
ME
Department of Biochemistry and Molecular Biology, University of Louisville Health Sciences Center, 501 South Preston St., Room 315, Louisville, KY 40292, USA.
Stearman
Randi P
RP
Smith
Gregory E
GE
Darling
Douglas S
DS
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D051548:Histone Acetyltransferases; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D009097:Multienzyme Complexes; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D014157:Transcription Factors / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000071799:Zinc Finger E-box-Binding Homeobox 1; D016335:Zinc Fingers; D001619:beta-N-Acetylhexosaminidases
D018398:Homeodomain Proteins; D009097:Multienzyme Complexes; D009687:Nuclear Proteins; D014157:Transcription Factors; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D051548:Histone Acetyltransferases; C110628:hexosaminidase C; D000118:Acetylglucosaminidase; D001619:beta-N-Acetylhexosaminidases