Complex regulation of somatic hypermutation by cis-acting sequences in the endogenous IgH gene in hybridoma cells

Diana Ronai, Maria Dolores Iglesias-Ussel, Manxia Fan, Marc J. Shulman, Matthew D. Scharff

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

To create high-affinity antibodies, B cells target a high rate of somatic hypermutation (SHM) to the Ig variable-region genes that encode the antigen-binding site. This mutational process requires transcription and is triggered by activation-induced cytidine deaminase (AID), which converts deoxycytidine to deoxyuridine. Mistargeting of AID to non-Ig genes is thought to result in the malignant transformation of B cells, but the mechanism responsible for targeting SHM to certain DNA regions and not to others is largely unknown. Cis-acting elements have been proposed to play a role in directing the hypermutation machinery, but the motifs required for targeting SHM have been difficult to identify because many of the candidate elements, such as promoters or enhancers, are also required for transcription of Ig genes. Here we describe a system in cultured hybridoma cells in which transcription of the endogenous heavy-chain Ig gene continues in the absence of the core intronic enhancer (Eμ) and its flanking matrix attachment regions (MARs). When AID is expressed in these cells, SHM occurred at the WT frequency even when Eμ and the MARs were absent together. Interestingly, SHM occurred at less than the WT frequency when Eμ or the MARs were individually absent. Our results suggest that these intronic regulatory elements can exert a complex influence on SHM that is separable from their role in regulating transcription.

Original languageEnglish (US)
Pages (from-to)11829-11834
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number33
DOIs
StatePublished - Aug 16 2005

Fingerprint

Matrix Attachment Regions
Hybridomas
Immunoglobulin Genes
B-Lymphocytes
Immunoglobulin Variable Region
Genes
Deoxyuridine
Deoxycytidine
Antibody Affinity
Cultured Cells
Binding Sites
Antigens
DNA
AICDA (activation-induced cytidine deaminase)

Keywords

  • cis-acting elements
  • Enhancer
  • Matrix attachment region

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Complex regulation of somatic hypermutation by cis-acting sequences in the endogenous IgH gene in hybridoma cells. / Ronai, Diana; Iglesias-Ussel, Maria Dolores; Fan, Manxia; Shulman, Marc J.; Scharff, Matthew D.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 102, No. 33, 16.08.2005, p. 11829-11834.

Research output: Contribution to journalArticle

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