IgTCR基因重排原理及多样性.pptxVIP

会计学1IgTCR基因重排原理及多样性 Ig gene rearrangement in antibody producing cells - Southern blot analysis non lymphoid cell : V,C in the distant fragments mature B cell : V,C in the same fragments V region of light chain : V gene segment (VL protein) (VL gene) J gene segment (JL gene) V region of heavy chain : V gene segment (VH protein) (VH gene) D gene segment (DH gene) J gene segment (JH gene)第1页/共26页 第2页/共26页 第3页/共26页 - The numbers of functional gene segments for the V region of human heavy and light chains ′ pseudogenes ′ : not functional proteinThe germline organization of the Ig genes gene family 第4页/共26页 第5页/共26页 第6页/共26页 Rearrangement of V,D,J gene segments - DNA rearrangements are guided by conserved (recombination signal sequence, RSS) ? heptamer : contiguous with the coding sequence ? space : 12 or 23 nucleotides long ? nonamer : second conserved sequence - coding sequence-heptamer-space-nonamer (12/23) - 12/23 rule - exception : D-D joining (fusion) in human ≈ 5% ? looping out and deletion : more common ? no deletion 第7页/共26页 第8页/共26页 第9页/共26页 第10页/共26页 第11页/共26页 Enzymatic steps in the gene rearrangement - RAG (Recombination Activating Genes RAG-1,RAG-2) protein complexes bind to RSSs ? make single-strand DNA breaks at sites 5′ of each RSS - RAG are only expressed in developing lymphocytes - Other ubiquitously expressed DNA-modifying proteins ? DNA ligase IV : joins the processed ends together with XRCC4 ? DNA-dependent protein kinase (DNA-PK) ? K