Functional interrogation of an odorant receptor locus reveals multiple axes of transcriptional regulation

PLoS Biol. 2013;11(5):e1001568. doi: 10.1371/journal.pbio.1001568. Epub 2013 May 21.

Abstract

The odorant receptor (OR) genes constitute the largest mammalian gene family and are expressed in a monogenic and monoallelic fashion, through an unknown mechanism that likely exploits positive and negative regulation. We devised a genetic strategy in mice to examine OR selection by determining the transcriptional activity of an exogenous promoter homologously integrated into an OR locus. Using the tetracycline-dependent transactivator responsive promoter (tet(o)), we observed that the OR locus imposes spatial and temporal constraints on tet(o)-driven transcription. Conditional expression experiments reveal a developmental change in the permissiveness of the locus. Further, expression of an OR transgene that suppresses endogenous ORs similarly represses the OR-integrated tet(o). Neurons homozygous for the tet(o)-modified allele demonstrate predominantly monoallelic expression, despite their potential to express both copies. These data reveal multiple axes of regulation, and support a model of initiation of OR choice limited by nonpermissive chromatin and maintained by repression of nonselected alleles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Animals
  • Gene Expression Regulation
  • Genes, Reporter
  • Mice
  • Mice, Inbred C57BL
  • Promoter Regions, Genetic
  • Receptors, Odorant / genetics*
  • Tetracycline
  • Transcriptional Activation*
  • Transgenes

Substances

  • Receptors, Odorant
  • Tetracycline

Grants and funding

This work was supported by Qatar National Research Fund (QNRF) grant NPRP 09-1011-3-260, and by Biomedical Research Program (BMRP) funds at Weill Cornell Medical College in Qatar, from the Qatar Foundation. AF was supported by a Marie Curie International Reintegration Grant and the ‘Amorcage de jeunes equipes’ program of the Fondation pour la Recherche Medical. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.