Homeodomain binding motifs modulate the probability of odorant receptor gene choice in transgenic mice.

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Version: Final published version
Serval ID
serval:BIB_A74DE94AE004
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Homeodomain binding motifs modulate the probability of odorant receptor gene choice in transgenic mice.
Journal
Molecular and Cellular Neuroscience
Author(s)
Vassalli A., Feinstein P., Mombaerts P.
ISSN
1095-9327 (Electronic)
ISSN-L
1044-7431
Publication state
Published
Issued date
2011
Volume
46
Number
2
Pages
381-396
Language
english
Abstract
Odorant receptor (OR) genes constitute with 1200 members the largest gene family in the mouse genome. A mature olfactory sensory neuron (OSN) is thought to express just one OR gene, and from one allele. The cell bodies of OSNs that express a given OR gene display a mosaic pattern within a particular region of the main olfactory epithelium. The mechanisms and cis-acting DNA elements that regulate the expression of one OR gene per OSN - OR gene choice - remain poorly understood. Here, we describe a reporter assay to identify minimal promoters for OR genes in transgenic mice, which are produced by the conventional method of pronuclear injection of DNA. The promoter transgenes are devoid of an OR coding sequence, and instead drive expression of the axonal marker tau-β-galactosidase. For four mouse OR genes (M71, M72, MOR23, and P3) and one human OR gene (hM72), a mosaic, OSN-specific pattern of reporter expression can be obtained in transgenic mice with contiguous DNA segments of only ~300 bp that are centered around the transcription start site (TSS). The ~150bp region upstream of the TSS contains three conserved sequence motifs, including homeodomain (HD) binding sites. Such HD binding sites are also present in the H and P elements, DNA sequences that are known to strongly influence OR gene expression. When a 19mer encompassing a HD binding site from the P element is multimerized nine times and added upstream of a MOR23 minigene that contains the MOR23 coding region, we observe a dramatic increase in the number of transgene-expressing founders and lines and in the number of labeled OSNs. By contrast, a nine times multimerized 19mer with a mutant HD binding site does not have these effects. We hypothesize that HD binding sites in the H and P elements and in OR promoters modulate the probability of OR gene choice.
Keywords
Amino Acid Motifs/genetics, Animals, Base Sequence, Conserved Sequence, Gene Expression Regulation/genetics, Humans, Immunohistochemistry, Mice, Mice, Transgenic, Microscopy, Confocal, Molecular Sequence Data, Olfactory Receptor Neurons/metabolism, Promoter Regions, Genetic/genetics, Receptors, Odorant/genetics, Reverse Transcriptase Polymerase Chain Reaction
Pubmed
Web of science
Open Access
Yes
Create date
28/03/2011 10:39
Last modification date
20/08/2019 15:12
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