Identification Of New Binding-partners Of Nogo And Function Analysis Of Nogo And BEX1

In present study, the human brain cDNA library was screened with Nogo-66 by using yeast two-hybrid system, BEX1, clusterin and some other gene fragments have been identified. Identification of interaction between Nogo and these candidate molecules were demonstrated by immunoprecipitaion.: Nogo-B/ Nogo-A could bind to BEX1 and p75NTR was also detected in the binding compound, which meant that Nogo could bind with p75NTR and BEX1. And from the other results , it could been found Nogo-B could bind with both clusterin and Necdin .Flow Cytometry results showed Nogo-B could induce obvious apoptosis in 293T, while Nogo-A not. And over-expression of BEX1,clusterin and Necdin all can de- crease apoptosis induced by Nogo-B, but BEX1 could sharply decrease the Nogo-B induced apoptosis. Hoechest nuclei staining results also supported BEX1could relieve Nogo-B induced apoptosis.Fluro microscopy observation and Hoechest staining showed that Nogo-B could make BEX1 translocate from nuclei to cytosome. And most of the translocation to cytosome cells manifested apoptosis performance.Fluroscence bearing cells calculation and western Blot showed Nogo-A and Nogo-B both could decrease BEX1 expression level, especially Nogo-A could greatly decrease BEX1 expression(P<0.01). RT-PCR analysis showed Nogo-A could obviously decrease BEX1 mRNA level, but Nogo-B seemed not impact BEX1 mRNA transcription obviously, which suggested that Nogo-A could change BEX1 expression level based on mRNA regulation.Totally speaking,except apoptosis induced Nogo-B were reported by Yutsudo in 2003, while denied by Schwab in 2003,the other results in the above totally have not been reported .