Replacement of Imu-Cmu intron by NeoR gene alters Imu germ-line expression but has no effect on V(D)J recombination - PubMed

Replacement of Imu-Cmu intron by NeoR gene alters Imu germ-line expression but has no effect on V(D)J recombination

Dania Haddad et al. Mol Immunol. 2010 Feb.

Abstract

The NeoR gene has often been used to unravel the mechanisms underlying long-range interactions between promoters and enhancers during V(D)J assembly and class switch recombination (CSR) in the immunoglobulin heavy chain (IgH) locus. This approach led to the notion that CSR is regulated through competition of germ-line (GL) promoters for activities displayed by the 3' regulatory region (3'RR). This polarized long-range effect of the 3'RR is disturbed upon insertion of NeoR gene in the IgH constant (C(H)) region, where only GL transcription derived from upstream GL promoters is impaired. In the context of V(D)J recombination, replacement of Emu enhancer or Emu core enhancer (cEmu) by NeoR gene fully blocked V(D)J recombination and mu0 GL transcription which originates 5' of DQ52 and severely diminished Imu GL transcription derived from Emu/Imu promoter, suggesting a critical role for cEmu in the regulation of V(D)J recombination and of mu0 and Imu expression. Here we focus on the effect of NeoR gene on mu0 and Imu GL transcription in a mouse line in which the Imu-Cmu intron was replaced by a NeoR gene in the sense-orientation. B cell development was characterized by a marked but incomplete block at the pro-B cell stage. However, V(D)J recombination was unaffected in sorted pro-B and pre-B cells excluding an interference with the accessibility control function of Emu. mu0 GL transcription initiation was relatively normal but the maturation step seemed to be affected most likely through premature termination at NeoR polyadenylation sites. In contrast, Imu transcription initiation was impaired suggesting an interference of NeoR gene with the IgH enhancers that control Imu expression. Surprisingly, in stark contrast with the NeoR effect in the C(H) region, LPS-induced NeoR expression restored Imu transcript levels to normal. The data suggest that Emu enhancer may be the master control element that counteracts the down-regulatory "Neo effect" on Imu expression upon LPS stimulation. More importantly, they reveal a complex and developmentally regulated interplay between IgH enhancers in the control of Imu expression.

Similar articles

• Elucidation of IgH intronic enhancer functions via germ-line deletion.

  Perlot T, Alt FW, Bassing CH, Suh H, Pinaud E. Perlot T, et al. Proc Natl Acad Sci U S A. 2005 Oct 4;102(40):14362-7. doi: 10.1073/pnas.0507090102. Epub 2005 Sep 26. Proc Natl Acad Sci U S A. 2005. PMID: 16186486 Free PMC article.

• Insertion of an imprinted insulator into the IgH locus reveals developmentally regulated, transcription-dependent control of V(D)J recombination.

  Puget N, Hirasawa R, Hu NS, Laviolette-Malirat N, Feil R, Khamlichi AA. Puget N, et al. Mol Cell Biol. 2015 Feb;35(3):529-43. doi: 10.1128/MCB.00235-14. Epub 2014 Nov 17. Mol Cell Biol. 2015. PMID: 25403489 Free PMC article.

• Mechanism and control of V(D)J recombination versus class switch recombination: similarities and differences.

  Dudley DD, Chaudhuri J, Bassing CH, Alt FW. Dudley DD, et al. Adv Immunol. 2005;86:43-112. doi: 10.1016/S0065-2776(04)86002-4. Adv Immunol. 2005. PMID: 15705419 Review.

• The IgH locus 3' regulatory region: pulling the strings from behind.

  Pinaud E, Marquet M, Fiancette R, Péron S, Vincent-Fabert C, Denizot Y, Cogné M. Pinaud E, et al. Adv Immunol. 2011;110:27-70. doi: 10.1016/B978-0-12-387663-8.00002-8. Adv Immunol. 2011. PMID: 21762815 Review.

Cited by

• Sense transcription through the S region is essential for immunoglobulin class switch recombination.

  Haddad D, Oruc Z, Puget N, Laviolette-Malirat N, Philippe M, Carrion C, Le Bert M, Khamlichi AA. Haddad D, et al. EMBO J. 2011 Apr 20;30(8):1608-20. doi: 10.1038/emboj.2011.56. Epub 2011 Mar 4. EMBO J. 2011. PMID: 21378751 Free PMC article.

• AIDing Chromatin and Transcription-Coupled Orchestration of Immunoglobulin Class-Switch Recombination.

  Vaidyanathan B, Yen WF, Pucella JN, Chaudhuri J. Vaidyanathan B, et al. Front Immunol. 2014 Mar 28;5:120. doi: 10.3389/fimmu.2014.00120. eCollection 2014. Front Immunol. 2014. PMID: 24734031 Free PMC article. Review.

• AID targeting: old mysteries and new challenges.

  Chandra V, Bortnick A, Murre C. Chandra V, et al. Trends Immunol. 2015 Sep;36(9):527-35. doi: 10.1016/j.it.2015.07.003. Epub 2015 Aug 4. Trends Immunol. 2015. PMID: 26254147 Free PMC article. Review.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Elsevier Science
  • Ovid Technologies, Inc.

• Molecular Biology Databases

  • Mouse Genome Informatics (MGI)