vimentin野生型和e151k突变体稳定表达晶状体细胞株的建立细胞生物学专业论文.docxVIP

ABSTACTVimentin ABSTACT Vimentin is a type III intermediate filament protein that is expressed in endothelial and other mesenchymal cells．In the eye lens epithelial cells， vimentin is abundantly expressed with important functions．Structurally， vimentin has three domains．The central rod domain contains仅helix connecting both the N--terminal head domain and the C·-terminal tail domain．As a maj or cytoskeletal protein，vimentin in involved in connecting the nucleus and membrane as well as mediating signal transduction．The normal function of vimentin influences differentiation of lens cells，and assures cell movement since it is part of the pseudopodia moving machinery．The function of vimentin’is affected by phosphorylation in which the protein kinase C is known to act on some of the phosphorylation sites．In addition，vimentin is highly expressed in cancer cells that acquire invasiveness．The normal function of the lens is closely related to vimentin function since mutations in vimentin gene cause its assembly disorder．A classic example is G5 96A change in vimentin which codes for exon 1 with the production of the mutant protein E 1 5 1 K．This mutant protein leads to vimentin assembly disorder， causing cataract formation．Since the substrate binding with SUMO generally has the same motif qSCxE／D，where、壬，is a hydrophobic amino III 万方数据 acid，K acid，K is target lysine，X is any amino acid，SO E 1 5 1 K mutation will produce a very conservative SUMO sites．We speculate that the E 1 5 1 K mutation may lead to sumoylation of the vimentin monomers，causing absence of vimentin assembly and resulting in cataractogenesis．To prove this hypothesis，we cloned the vimentin cDNA from mouse lens epithelial cells and conducted in vitro mutagenesis to generate the wild type and mutant E 1 5 1 K vimentin expression constructs．Then，we transfected these constructs into mouse lens epithelial cells，0【-TN4—1 and cultured the transfected cells in the presence of the screening drug，G4 1 8(600 ug／m1)． A