Identification of ubiquitin-interacting proteins in purified polyglutamine aggregates.
Nuclear aggregates of enhanced green fluorescent protein and nuclear localization signal-fused truncated N-terminal huntingtin containing 150 repeats of glutamine residue were purified from ecdysine-inducible mutant neuro2A cell line by sequential extraction of nuclear soluble proteins. To analyze the aggregate-interacting proteins, we subjected the nuclear aggregates to high performance liquid chromatography-mass spectrometry analysis. The resulting data revealed the presence of three new putative aggregate-interacting proteins: ubiquilin 1, ubiquilin 2 and Tollip. These proteins also associated with neuronal intranuclear inclusions in a mouse model of Huntington disease (HD). These aggregate-interacting proteins contain ubiquitin-interacting motifs, suggesting that they are recruited to the aggregates where they may lose their normal function.
Research Resource Center, RIKEN Brain Science Institute, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
Hiroshi
H
Doi
Kenichi
K
Mitsui
Masaru
M
Kurosawa
Yoko
Y
Machida
Yoshiyuki
Y
Kuroiwa
Nobuyuki
N
Nukina
D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D005091:Exons; D005973:Glutamine / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D025801:Ubiquitin / Q000378:metabolism*
D008164:Luminescent Proteins; D010455:Peptides; D011994:Recombinant Proteins; D025801:Ubiquitin; D005973:Glutamine; D049452:Green Fluorescent Proteins; C097188:polyglutamine