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Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing
Published on: March 15, 2019
The mouse immunoglobulin heavy chain V-D intergenic sequence contains insulators that may regulate ordered V(D)J
Karen Featherstone1, Andrew L Wood1, Adam J Bowen1
1Laboratory of Chromatin and Gene Expression, Babraham Institute, Babraham Research Campus, Cambridge CB22 3AT, United Kingdom.
The Journal of Biological Chemistry
|January 27, 2010
Summary
Researchers identified novel regulatory elements controlling ordered immunoglobulin heavy chain V(D)J recombination. These elements, including enhancer-blocking insulators, ensure correct antibody gene assembly by regulating chromatin accessibility.
Area of Science:
- Immunology
- Molecular Biology
- Genetics
Background:
- Immunoglobulin heavy chain (Igh) V(D)J recombination is an ordered process crucial for antibody assembly.
- Differential chromatin accessibility controls the D to J and V to DJ recombination steps.
- The intronic enhancer Emu is known to regulate D to J recombination, but other cis-acting elements remain unidentified.
Purpose of the Study:
- To identify cis-acting regulatory elements controlling differential chromatin opening in the mouse Igh V-D intergenic region.
- To understand the regulation of ordered V(D)J recombination and antibody gene assembly.
Main Methods:
- Sequencing and analysis of a 96-kb V-D intergenic region in the mouse Igh.
- Assessing antisense transcription patterns during lymphocyte development.
- Identifying DNase I-hypersensitive sites (HSs) and analyzing their function in vivo.
- Investigating CTCF binding and enhancer-blocking activity of identified HSs.
Main Results:
- Emu-dependent D antisense transcription extends over 30 kb into the V-D region in B and T cells.
- This transcription is actively prevented from reaching the V genes by two conserved HSs near the D region.
- These HSs bind CTCF in vivo and exhibit enhancer-blocking activity, acting as insulators.
- One HS upstream of the D gene locally regulates D genes.
Conclusions:
- The identified HSs function as enhancer-blocking insulators, preventing premature chromatin opening into the V region.
- These elements are proposed to control ordered V(D)J recombination by regulating differential chromatin accessibility.
- This discovery provides the first identified elements that may govern the precise order of V(D)J recombination for correct antibody gene assembly.
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