Regions downstream from the WW domain of dystrophin are important for binding to postsynaptic densities in the brain.
In order to investigate the mechanism of dystrophin localization in the central nervous system (CNS), we generated adenovirus vectors that contained minidystrophin or truncated minidystrophin cDNA. We infected a primary neuronal culture derived from mdx mouse hippocampus with these viruses. Minidystrophin was observed along the plasma membrane as punctate dots or very short segments. In double immunofluorescence staining with anti-dystrophin and anti-postsynaptic density-95 antibodies, we observed that these proteins entirely colocalized. On the other hand, the truncated minidystrophin, which has deleted WW, cysteine-rich and C-terminal domains, was homogenously expressed in cytoplasm, neurites and axons. These findings suggest that a binding site to postsynaptic densities exists in the region extending from the WW domain to the C-terminal domain of dystrophin and that this site is necessary for binding to membrane.
Sakamoto
Tetsuro
T
Department of Neurology, Kumamoto University Graduate School of Medical Sciences, 1-1-1 Honjo, Kumamoto, Kumamoto 860-0811, Japan. sakatetsu@fc.kuh.kumamoto-u.ac.jp <sakatetsu@fc.kuh.kumamoto-u.ac.jp>
Arima
Toshiyuki
T
Ishizaki
Masatoshi
M
Kawano
Ryoko
R
Koide
Tatsuya
T
Uchida
Yuji
Y
Yamashita
Satoshi
S
Kimura
En
E
Hirano
Teruyuki
T
Maeda
Yasushi
Y
Uchino
Makoto
M
D000256:Adenoviridae / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D019556:COS Cells; D018929:Cell Culture Techniques; D002460:Cell Line; D002522:Chlorocebus aethiops; D016189:Dystrophin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D008969:Molecular Sequence Data; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D013569:Synapses / Q000378:metabolism*; D014162:Transfection
D016189:Dystrophin; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins