Integrated biochemical and computational approach identifies BCL6 direct target genes controlling multiple pathways in normal germinal center B cells

Blood. 2010 Feb 4;115(5):975-84. doi: 10.1182/blood-2009-06-227017. Epub 2009 Dec 3.

Abstract

BCL6 is a transcriptional repressor required for mature B-cell germinal center (GC) formation and implicated in lymphomagenesis. BCL6's physiologic function is only partially known because the complete set of its targets in GC B cells has not been identified. To address this issue, we used an integrated biochemical-computational-functional approach to identify BCL6 direct targets in normal GC B cells. This approach includes (1) identification of BCL6-bound promoters by genome-wide chromatin immunoprecipitation, (2) inference of transcriptional relationships by the use of a regulatory network reverse engineering approach (ARACNe), and (3) validation of physiologic relevance of the candidate targets down-regulated in GC B cells. Our approach demonstrated that a large set of promoters (> 4000) is physically bound by BCL6 but that only a fraction of them is repressed in GC B cells. This set of 1207 targets identifies several cellular functions directly controlled by BCL6 during GC development, including activation, survival, DNA-damage response, cell cycle arrest, cytokine signaling, Toll-like receptor signaling, and differentiation. These results define a broad role of BCL6 in preventing centroblasts from responding to signals leading to exit from the GC before they complete the phase of proliferative expansion and of antibody affinity maturation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes / cytology
  • B-Lymphocytes / metabolism*
  • Binding Sites / genetics
  • Cell Line
  • Cell Line, Tumor
  • Cells, Cultured
  • Chromatin Immunoprecipitation
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Profiling
  • Gene Regulatory Networks
  • Genome-Wide Association Study / methods
  • Germinal Center / cytology
  • Germinal Center / metabolism*
  • Humans
  • Immunoblotting
  • Lymphocyte Activation
  • Oligonucleotide Array Sequence Analysis
  • Promoter Regions, Genetic / genetics*
  • Protein Binding
  • Proto-Oncogene Proteins c-bcl-6
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / genetics*
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism

Substances

  • BCL6 protein, human
  • DNA-Binding Proteins
  • Proto-Oncogene Proteins c-bcl-6

Associated data

  • GEO/GSE12195
  • GEO/GSE2350