中科院细胞与分子免疫学课程课件Chapter.pptVIP

* D J 7 12 9 V4 7 23 9 V4 and DJ in opposite transcriptional orientations D J 7 12 9 V4 7 23 9 1. D J 7 12 9 V4 7 23 9 3. D J 7 12 9 V4 7 23 9 2. D J 7 12 9 V4 7 23 9 4. Non-deletional recombination * D J 7 12 9 V4 7 23 9 1. D J V4 7 12 9 7 23 9 3. V to DJ ligation - coding joint formation D J 7 12 9 V4 7 23 9 2. Heptamer ligation - signal joint formation D J V4 7 12 9 7 23 9 Fully recombined VDJ regions in same transcriptional orientation No DNA is deleted 4. * 7 D 12 9 J 6.Generation of Diversity of the B and T Cell Repertoires Junctional diversity: P nucleotide additions 7 V 23 9 D 7 12 9 J V 7 23 9 TC CACAGTG AG GTGTCAC AT GTGACAC TA CACTGTG The recombinase complex makes single stranded nicks at random sites close to the ends of the V and D region DNA. 7 D 12 9 J 7 V 23 9 CACAGTG GTGTCAC GTGACAC CACTGTG TC AG AT TA D J V TC AG AT TA U U The 2nd strand is cleaved and hairpins form between the complementary bases at ends of the V and D region. * V2 V3 V4 V8 V7 V6 V5 V9 7 23 9 CACAGTG GTGTCAC 7 12 9 GTGACAC CACTGTG V TC AG U D J AT TA U Heptamers are ligated by DNA ligase IV V and D regions juxtaposed V TC AG U D J AT TA U * V TC AG U D J AT TA U Endonuclease cleaves single strand at random sites in V and D segment V TC~GA AG D J AT TA~TA The nucleotides that flip out, become part of the complementary DNA strand 6.1 Generation of the palindromic sequence In terms of G to C and T to A pairing, the ‘new’ nucleotides are palindromic. The nucleotides GA and TA were not in the genomic sequence and introduce diversity of sequence at the V to D join. V TC AG U D J AT TA U Regions to be joined are juxtaposed The nicked strand ‘flips’ out * 6.2 Junctional Diversity?–?N nucleotide additions V TC~GA AG D J AT TA~TA Terminal deoxynucleotidyl transferase (TdT) adds nucleotides randomly to the P nucleotide ends of the single-stranded V and D segment DNA CACTCCTTA TTCTTGCAA V TC~GA AG D J AT TA~TA CACACCTTA TTCTTGCAA Complementary bases anneal V D J DNA polymerases fill in