A novel interaction between ATOH8 and PPP3CB.
ATOH8 is a bHLH transcription factor playing roles in a variety of developmental processes such as neurogenesis, differentiation of pancreatic precursor cells, development of kidney and muscle, and differentiation of endothelial cells. PPP3CB belongs to the catalytic subunit of the serine/threonine phosphatase, calcineurin, which can dephosphorylate its substrate proteins to regulate their physiological activities. In our study, we demonstrated that ATOH8 interacts with PPP3CB in vitro with different approaches. We show that the conserved catalytic domain of PPP3CB interacts with both the N-terminus and the bHLH domain of ATOH8. Although the interaction domain of PPP3CB is conserved among all isoforms of calcineurin A, ATOH8 selectively interacts with PPP3CB instead of PPP3CA, probably due to the unique proline-rich region present in the N-terminus of PPP3CB, which controls the specificity of its interaction partners. Furthermore, we show that inhibition of the interaction with calcineurin inhibitor, cyclosporin A (CsA), leads to the retention of ATOH8 to the cytoplasm, suggesting that the interaction renders nuclear localization of ATOH8 which may be critical to control its activity as transcription factor.
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany.|Department of Craniofacial Development and Stem Cell Biology, King's College London, SE19RT, London, UK.
Jingchen
J
Chen
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany.
Ajeesh
A
Balakrishnan-Renuka
Department of Neurology, University Hospital Essen, 45122, Essen, Germany.
Nina
N
Hagemann
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany.|Department of Cytology, Ruhr-University Bochum, 44780, Bochum, Germany.
Carsten
C
Theiss
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany.
Verena
V
Chankiewitz
Department of Genetics, Fudan University, Shanghai, People's Republic of China.
Jinzhong
J
Chen
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany.
Qin
Q
Pu
Department of Biomedical Science & Centre for Membrane Interactions and Dynamics (CMIAD), University of Sheffield, S10 2TN , Sheffield, UK.
Kai S
KS
Erdmann
Department of Anatomy and Molecular Embryology, Medizinische Fakultät, Ruhr-Universität Bochum, Abt. f. Anatomie und Molekulare Embryologie, Geb. MA, 5/158, 44780, Bochum, Germany. Beate.Brand-Saberi@rub.de.
Beate
B
Brand-Saberi
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000737:chemistry* / Q000235:genetics; D019556:COS Cells; D019703:Calcineurin / Q000737:chemistry* / Q000235:genetics; D065095:Calcineurin Inhibitors / Q000737:chemistry*; D020013:Calcium Signaling / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002522:Chlorocebus aethiops; D016572:Cyclosporine / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
D051792:Basic Helix-Loop-Helix Proteins; D019703:Calcineurin; D065095:Calcineurin Inhibitors; D016572:Cyclosporine; C486727:ATOH8 protein, human; C525673:PPP3CA protein, human; C525674:PPP3CB protein, human
ATOH8; Calcineurin; Cyclosporin A; PPP3CB