title: [Molecular mechanisms of two novel mutations of factor XIII gene resulting in hereditary coagulation deficiency].
abstract: OBJECTIVE
  To investigate the mechanisms of two novel missense mutations of factor XIIIA subunit gene (Arg77-->Cys,Ser413-->Trp) in the pathogenesis of hereditary factor XIII deficiency.
  
  
  METHODS
  Site-directed mutagenesis was conducted to obtain 2 mutant human XIII A recombinant plasmids, mut-PCI/FXIIIA. Normal wild type factor XIII A recombinant plasmid, wt-PCI/FXIIIA, and mut-PCI/FXIIIA, were transfected into cultured COS7 cells line, renal fibroid cell of African green monkey using Superfect reagent respectively, The expression levels of DNA, RNA and protein of human factor XIII, both wild type and mutant, were detected by PCR, RT-PCR and Western blotting. Pulse-chase experiment was used to look into the changing of factor XIII A in the cytoplasm. Factor XIIIA activity was assayed by Biotin-pentylamine incorporation technique.
  
  
  RESULTS
  The mRNA levels of the two mutants in transfected cells were similar to that of the wild type factor XIIIA. But the amount of mutant factor XIIIA protein and its activity in cells decreased markedly, even disappeared. Pulse-chase experiment revealed that at the two mutants existed chase time 0.5 h and 1 h considerable amounts in cells and then disappeared rapidly later.
  
  
  CONCLUSIONS
  The 2 mutations of the factor XIIIA cause the instability, degradation, and rapid disappearance of FXIIIA in cytoplasm, thus resulting in hereditary factor XIII deficiency.
authors.affiliation: Shanghai Institute of Hematology, Ruijin Hospital Affiliated to Shanghai Second Medical University, Shanghai 200025, China.
authors.forename: Bao-hua
authors.initials: BH
authors.lastname: Duan
authors.forename: Hong-li
authors.initials: HL
authors.lastname: Wang
authors.forename: Xue-feng
authors.initials: XF
authors.lastname: Wang
authors.forename: Yi-qun
authors.initials: YQ
authors.lastname: Hu
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Chu
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Yin
authors.forename: Xue-mei
authors.initials: XM
authors.lastname: Guo
authors.forename: Qi-hua
authors.initials: QH
authors.lastname: Fu
authors.forename: Wen-man
authors.initials: WM
authors.lastname: Wu
authors.forename: Qiu-lan
authors.initials: QL
authors.lastname: Ding
authors.forename: Yi
authors.initials: Y
authors.lastname: Fang
authors.forename: Wen-bin
authors.initials: WB
authors.lastname: Wang
authors.forename: Rong-fu
authors.initials: RF
authors.lastname: Zhou
authors.forename: Wen-ying
authors.initials: WY
authors.lastname: Kang
authors.forename: Shuang
authors.initials: S
authors.lastname: Xie
authors.forename: Zhen-yi
authors.initials: ZY
authors.lastname: Wang
mesh_terms: D000818:Animals; D019556:COS Cells; D005176:Factor XIII / Q000737:chemistry / Q000235:genetics*; D005177:Factor XIII Deficiency / Q000235:genetics*; D006801:Humans; D009154:Mutation; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D012333:RNA, Messenger; D005176:Factor XIII