A cis-acting diversification activator both necessary and sufficient for AID-mediated hypermutation

PLoS Genet. 2009 Jan;5(1):e1000332. doi: 10.1371/journal.pgen.1000332. Epub 2009 Jan 9.

Abstract

Hypermutation of the immunoglobulin (Ig) genes requires Activation Induced cytidine Deaminase (AID) and transcription, but it remains unclear why other transcribed genes of B cells do not mutate. We describe a reporter transgene crippled by hypermutation when inserted into or near the Ig light chain (IgL) locus of the DT40 B cell line yet stably expressed when inserted into other chromosomal positions. Step-wise deletions of the IgL locus revealed that a sequence extending for 9.8 kilobases downstream of the IgL transcription start site confers the hypermutation activity. This sequence, named DIVAC for diversification activator, efficiently activates hypermutation when inserted at non-Ig loci. The results significantly extend previously reported findings on AID-mediated gene diversification. They show by both deletion and insertion analyses that cis-acting sequences predispose neighboring transcription units to hypermutation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes
  • Chickens
  • Chromosome Mapping
  • Cytidine Deaminase / metabolism*
  • Flow Cytometry
  • Gene Deletion
  • Gene Expression Regulation
  • Genes, Immunoglobulin Light Chain*
  • Genes, Reporter
  • Genetic Variation
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Mutation
  • Regulatory Sequences, Nucleic Acid*
  • Sequence Analysis, DNA
  • Somatic Hypermutation, Immunoglobulin*
  • Transfection

Substances

  • Green Fluorescent Proteins
  • AICDA (activation-induced cytidine deaminase)
  • Cytidine Deaminase