title: The TSC1 gene product hamartin interacts with NADE.
abstract: Hamartomatous brain lesions are a hallmark of brain pathology of tuberous sclerosis complex (TSC). To elucidate the mechanism of tumor development in the brain of TSC, we identified NADE (p75NTR-associated cell death executor) as an interactor for TSC1 gene product hamartin using a yeast two-hybrid system. In a pull-down assay, endogenous NADE was purified with the immobilized coiled-coil domain (CCD) of hamartin from the PC12h cell lysate. Immunofluorescence and immunoprecipitation confirmed the interaction of hamartin and NADE in cultured neurons and mouse brain lysate. Hamartin constitutively associated with NADE to prevent its proteasomal degradation. Suppression of hamartin with TSC1 small interfering RNA (siRNA) caused reduction of NADE and failed to lead to NGF-induced apoptosis in PC12h cells. These results indicate that hamartin binds to NADE to regulate neuronal cell function and loss of this association is likely to contribute to the brain pathology in TSC.
authors.affiliation: Department of Neurobiology, School of Life Sciences, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.
authors.forename: Sakiko
authors.initials: S
authors.lastname: Yasui
authors.forename: Kokoro
authors.initials: K
authors.lastname: Tsuzaki
authors.forename: Haruaki
authors.initials: H
authors.lastname: Ninomiya
authors.forename: Florin
authors.initials: F
authors.lastname: Floricel
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Asano
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Maki
authors.forename: Ayumi
authors.initials: A
authors.lastname: Takamura
authors.forename: Eiji
authors.initials: E
authors.lastname: Nanba
authors.forename: Katsumi
authors.initials: K
authors.lastname: Higaki
authors.forename: Kousaku
authors.initials: K
authors.lastname: Ohno
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019556:COS Cells; D002531:Cerebellum / Q000166:cytology; D002522:Chlorocebus aethiops; D006801:Humans; D007668:Kidney / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D020932:Nerve Growth Factor / Q000378:metabolism; D016716:PC12 Cells; D017434:Protein Structure, Tertiary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014402:Tuberous Sclerosis / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D000077004:Tuberous Sclerosis Complex 1 Protein; D025521:Tumor Suppressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D051017:Apoptosis Regulatory Proteins; C410727:BEX3 protein, human; D011993:Recombinant Fusion Proteins; C000624650:TSC1 protein, human; C000624651:Tsc1 protein, mouse; C000624652:Tsc1 protein, rat; D000077004:Tuberous Sclerosis Complex 1 Protein; D025521:Tumor Suppressor Proteins; D020932:Nerve Growth Factor