Immunoglobulin V regions diversify by gene conversion in DT40 cells
description
Transcript of Immunoglobulin V regions diversify by gene conversion in DT40 cells
Immunoglobulin V regions diversify by gene conversion in DT40 cells
Able to assay of sIgM+status by flow cytometry
V V V V V J C1
V V V V V* J C1
V
V
V*
Chromatin features of pseudogene donors
Histone acetylation
H3.3 variant
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1000
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10
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Forward scatter Forward scatter
s IG
M
s IG
M
GFP-LacI HIRA-LacI
Effects on diversification
Med
ian
sIg
M
loss
1.0 1.7 11.0 8.4
0
2
4
6
8
10
12
14
GFP-LacI HIRA Cl18 HIRA-LacI VP16-LacI
Long tract GC77%
Short tract GC3%
Point Mutation20%
Insertion0%
Deletion0%
GFP-LacI
71
Long tract GC66%
Short tract GC21%
Point Mutation11%
Insertion2%
Deletion0%
HIRA-LacI
137
VP16-LacI
Long-tract GC45%
Short-tract GC29%
Point Mutation13%
Deletion0%
Insertion13%
31
LacI-HP1Long tract GC
3%Short tract GC
17%
Point Mutation58%
Insertion8%
Deletion14%
36
Long tract GC77%
Short tract GC3%
Point Mutation20%
Insertion0%
Deletion0%
GFP-LacI
71
Long tract GC66%
Short tract GC21%
Point Mutation11%
Insertion2%
Deletion0%
HIRA-LacI
137
VP16-LacI
Long-tract GC45%
Short-tract GC29%
Point Mutation13%
Deletion0%
Insertion13%
31
LacI-HP1Long tract GC
3%Short tract GC
17%
Point Mutation58%
Insertion8%
Deletion14%
36
GFP-LacI-11 HIRA-LacI-19
Effect of HIRA on chromatin
Cell 1
Cell 2
nucleosomedeposition
Cell 1
Cell 2
~150 bp
MNase
VP16-LacI-10GFP-LacI-11
AcH4 IgG AcH4 IgG
V17
Ova
Enrichment: 4.5 12.4
GFP-LacI VP16-LacI
Effect of VP16 on chromatin