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Merck
  • Stage-specific binding profiles of cohesin in resting and activated B lymphocytes suggest a role for cohesin in immunoglobulin class switching and maturation.

Stage-specific binding profiles of cohesin in resting and activated B lymphocytes suggest a role for cohesin in immunoglobulin class switching and maturation.

PloS one (2014-11-07)
Gamze Günal-Sadık, Maciej Paszkowski-Rogacz, Kalaimathy Singaravelu, Andreas Beyer, Frank Buchholz, Rolf Jessberger
摘要

The immunoglobulin heavy chain locus (Igh) features higher-order chromosomal interactions to facilitate stage-specific assembly of the Ig molecule. Cohesin, a ring-like protein complex required for sister chromatid cohesion, shapes chromosome architecture and chromatin interactions important for transcriptional regulation and often acts together with CTCF. Cohesin is likely involved in B cell activation and Ig class switch recombination. Hence, binding profiles of cohesin in resting mature murine splenic B lymphocytes and at two stages after cell activation were elucidated by chromatin immunoprecipitation and deep sequencing. Comparative genomic analysis revealed cohesin extensively changes its binding to transcriptional control elements after 48 h of stimulation with LPS/IL-4. Cohesin was clearly underrepresented at switch regions regardless of their activation status, suggesting that switch regions need to be cohesin-poor. Specific binding changes of cohesin at B-cell specific gene loci Pax5 and Blimp-1 indicate new cohesin-dependent regulatory pathways. Together with conserved cohesin/CTCF sites at the Igh 3'RR, a prominent cohesin/CTCF binding site was revealed near the 3' end of Cα where PolII localizes to 3' enhancers. Our study shows that cohesin likely regulates B cell activation and maturation, including Ig class switching.

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Sigma-Aldrich
脂多糖 来源于肠沙门氏菌 肠道血清型, purified by phenol extraction
Sigma-Aldrich
抗CTCF抗体, serum, Upstate®
Sigma-Aldrich
Interleukin-4 from mouse, IL-4, recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
Sigma-Aldrich
IL-4 来源于小鼠, Animal-component free, recombinant, expressed in E. coli, ≥98% (SDS-PAGE), ≥98% (HPLC), suitable for cell culture
Sigma-Aldrich
(±)β-羟基异丁酸钠, ≥96.0%