Immunoglobulin switch μ sequence causes RNA polymerase II accumulation and reduces dA hypermutation

Deepa Rajagopal, Robert W. Maul, Amalendu Ghosh, Tirtha Chakraborty, Ahmed Amine Khamlichi, Ranjan Sen, Patricia J. Gearhart

Research output: Contribution to journalArticle

Abstract

Repetitive DNA sequences in the immunoglobulin switch μ region form RNA-containing secondary structures and undergo hypermutation by activation-induced deaminase (AID). To examine how DNA structure affects transcription and hypermutation, we mapped the position of RNA polymerase II molecules and mutations across a 5-kb region spanning the intronic enhancer to the constant μ gene. For RNA polymerase II, the distribution was determined by nuclear run-on and chromatin immunoprecipitation assays in B cells from uracil-DNA glycosylase (UNG)-deficient mice stimulated ex vivo. RNA polymerases were found at a high density in DNA flanking both sides of a 1-kb repetitive sequence that forms the core of the switch region. The pileup of polymerases was similar in unstimulated and stimulated cells from Ung- /- and Aid- /- Ung- /- mice but was absent in cells from mice with a deletion of the switch region. For mutations, DNA was sequenced from Ung- /- B cells stimulated in vivo. Surprisingly, mutations of A nucleotides, which are incorporated by DNA polymerase η, decreased 10-fold before the repetitive sequence, suggesting that the polymerase was less active in this region. We propose that altered DNA structure in the switch region pauses RNA polymerase II and limits access of DNA polymerase η during hypermutation.

Original languageEnglish (US)
Pages (from-to)1237-1244
Number of pages8
JournalJournal of Experimental Medicine
Volume206
Issue number6
DOIs
StatePublished - Jun 8 2009

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Rajagopal, D., Maul, R. W., Ghosh, A., Chakraborty, T., Khamlichi, A. A., Sen, R., & Gearhart, P. J. (2009). Immunoglobulin switch μ sequence causes RNA polymerase II accumulation and reduces dA hypermutation. Journal of Experimental Medicine, 206(6), 1237-1244. https://doi.org/10.1084/jem.20082514