Pro-apoptotic ASY/Nogo-B protein associates with ASYIP.
We have previously shown that ectopic expression of the ASY/Nogo-B gene induced apoptosis in various cancer cell lines. Nogo-A, a splice variant of the ASY, has been reported to have an inhibitory effect on neuronal regeneration in the central nervous system. To investigate the mechanism of ASY-induced apoptosis or inhibition of neuronal regeneration, we cloned a cDNA for the ASY-interacting protein from the human cDNA library using the yeast two-hybrid method, and obtained a cDNA we designated as ASYIP. The ASYIP protein contains two hydrophobic regions and a double lysine endoplasmic reticulum (ER) retrieval motif at its C-terminus, which was shown to be identical to RTN3, a reticulon family protein of unknown function. We showed that ASY and ASYIP proteins formed a complex also in human cells. Mutational analysis indicated that both of the hydrophobic regions of the ASYIP protein were required for the association. By immunofluorescence analysis, the ASYIP protein was shown to be co-localized with ASY in the ER. Characterization of the ASYIP gene may be very useful in clarifying the mechanism of ASY-induced apoptosis or Nogo-involved inhibition of neuronal regeneration in the central nervous system.
Qi
Bing
B
Institute of Virology, Wuhan University, Wuhan, Hubei, Peoples Republic of China.
Qi
Yipeng
Y
Watari
Akihiro
A
Yoshioka
Naohisa
N
Inoue
Hirokazu
H
Minemoto
Yuzuru
Y
Yamashita
Katsumi
K
Sasagawa
Toshiyuki
T
Yutsudo
Masuo
M
D000595:Amino Acid Sequence / Q000235:genetics; D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D009185:Myelin Proteins / Q000378:metabolism / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D000070798:Nogo Proteins; D014018:Tissue Distribution; D020798:Two-Hybrid System Techniques; D015003:Yeasts
D002352:Carrier Proteins; D008565:Membrane Proteins; D009185:Myelin Proteins; D009419:Nerve Tissue Proteins; D000070798:Nogo Proteins; C476198:RTN3 protein, human; C000603199:RTN4 protein, human