Equal transcription rates of productively and nonproductively rearranged immunoglobulin μ heavy chain alleles in a pro-B cell line

  1. Andrea B. Eberle1,
  2. Kai Herrmann2,
  3. Hans-Martin Jäck2 and
  4. Oliver Mühlemann1
  1. 1Institute of Cell Biology, University of Bern, 3012 Bern, Switzerland
  2. 2Division of Molecular Immunology, Department of Internal Medicine III, Nikolaus-Fiebiger-Center, University of Erlangen-Nürnberg, 91054 Erlangen, Germany

    Abstract

    During B cell maturation, immunoglobulin (Ig) genes frequently acquire premature translation-termination codons (PTCs) as a result of the somatic rearrangement of V, D, and J gene segments. However, it is essential for a B lymphocyte to produce only one kind of antibody and therefore to ensure that the heavy and light chain polypeptides are expressed exclusively from the corresponding functional alleles, whereas no protein is made from the nonproductively rearranged alleles. At the post-transcriptional level, a well-studied mRNA quality control mechanism, termed nonsense-mediated mRNA decay (NMD), recognizes and degrades PTC-containing mRNAs in a translation-dependent manner. In addition, transcriptional silencing of PTC-containing Ig-μ and Ig-γ heavy chain reporter genes was observed in HeLa cells. To investigate the silencing of nonproductively rearranged Ig genes in a more physiological context, we analyzed a monoclonal line of immortalized murine pro-B cells harboring one productively (PTC−) and one nonproductively (PTC+) rearranged Ig-μ heavy chain allele. We show that the steady-state abundance of PTC+ mRNA was ∼40-fold lower when compared to that of the PTC− mRNA. However, both the PTC+ and PTC− allele seemed to be equally well transcribed since the abundances of PTC+ and PTC− pre-mRNA were very similar and chromatin immunoprecipitations revealed comparable occupancy of RNA polymerase II and acetylated histone H3 on both alleles. Altogether, we found no evidence for transcriptional silencing of the PTC+ allele in this pro-B cell line; hence, the efficient down-regulation of the PTC+ Ig-μ mRNA results entirely from NMD.

    Keywords

    Footnotes

    • Reprint requests to: Oliver Mühlemann, Institute of Cell Biology, University of Bern, Baltzerstrasse 4, 3012 Bern, Switzerland; e-mail: oliver.muehlemann{at}izb.unibe.ch; fax: +41-31-631-4616.

    • Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.1516409.

      • Received December 15, 2008.
      • Accepted February 24, 2009.

    Articles citing this article

    • The Ig heavy chain protein but not its message controls early B cell development Proc. Natl. Acad. Sci. USA December 8, 2020 117: 31343-31352
    • A plasma cell differentiation quality control ablates B cell clones with biallelic Ig rearrangements and truncated Ig production JEM January 11, 2016 213: 109-122
    • Complete cis Exclusion upon Duplication of the E{mu} Enhancer at the Immunoglobulin Heavy Chain Locus Mol. Cell. Biol. July 1, 2015 35: 2231-2241
    • Cross Talk between Immunoglobulin Heavy-Chain Transcription and RNA Surveillance during B Cell Development Mol. Cell. Biol. January 1, 2012 32: 107-117
    • Pro-B cells sense productive immunoglobulin heavy chain rearrangement irrespective of polypeptide production Proc. Natl. Acad. Sci. USA June 28, 2011 108: 10644-10649
    • Multiple RNA Surveillance Mechanisms Cooperate to Reduce the Amount of Nonfunctional Ig{kappa} Transcripts J. Immunol. May 1, 2010 184: 5009-5017
    | Table of Contents

    This Article

    1. Published in Advance April 10, 2009, doi: 10.1261/rna.1516409 RNA 15: 1021-1028 Copyright © 2009 RNA Society

    Article Category

    Share

    Current Issue

    1. August 2026, 32 (8)
    1. Current Issue
    1. Alert me to new issues of RNA