The TSC1 gene product hamartin interacts with NADE

Sakiko Yasui; Kokoro Tsuzaki; Haruaki Ninomiya; Florin Floricel; Yasuo Asano; Hirotoshi Maki; Ayumi Takamura; Eiji Nanba; Katsumi Higaki; Kousaku Ohno

doi:10.1016/j.mcn.2007.02.007

The TSC1 gene product hamartin interacts with NADE

Mol Cell Neurosci. 2007 May;35(1):100-8. doi: 10.1016/j.mcn.2007.02.007. Epub 2007 Feb 12.

Authors

Sakiko Yasui¹, Kokoro Tsuzaki, Haruaki Ninomiya, Florin Floricel, Yasuo Asano, Hirotoshi Maki, Ayumi Takamura, Eiji Nanba, Katsumi Higaki, Kousaku Ohno

Affiliation

¹ Department of Neurobiology, School of Life Sciences, Tottori University Faculty of Medicine, Yonago 683-8503, Japan.

PMID: 17355907
DOI: 10.1016/j.mcn.2007.02.007

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / physiology
Apoptosis Regulatory Proteins / genetics
Apoptosis Regulatory Proteins / isolation & purification
Apoptosis Regulatory Proteins / metabolism*
COS Cells
Cerebellum / cytology
Chlorocebus aethiops
Humans
Kidney / cytology
Mice
Mice, Inbred C57BL
Nerve Growth Factor / metabolism
PC12 Cells
Protein Structure, Tertiary
Rats
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / isolation & purification
Recombinant Fusion Proteins / metabolism
Tuberous Sclerosis / metabolism*
Tuberous Sclerosis / pathology
Tuberous Sclerosis / physiopathology
Tuberous Sclerosis Complex 1 Protein
Tumor Suppressor Proteins / chemistry
Tumor Suppressor Proteins / genetics
Tumor Suppressor Proteins / metabolism*
Two-Hybrid System Techniques

Substances

Apoptosis Regulatory Proteins
BEX3 protein, human
Recombinant Fusion Proteins
TSC1 protein, human
Tsc1 protein, mouse
Tsc1 protein, rat
Tuberous Sclerosis Complex 1 Protein
Tumor Suppressor Proteins
Nerve Growth Factor