A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep

Abstract

Texel sheep are renowned for their exceptional meatiness. To identify the genes underlying this economically important feature, we performed a whole-genome scan in a Romanov × Texel F2 population. We mapped a quantitative trait locus with a major effect on muscle mass to chromosome 2 and subsequently fine-mapped it to a chromosome interval encompassing the myostatin (GDF8) gene. We herein demonstrate that the GDF8 allele of Texel sheep is characterized by a G to A transition in the 3′ UTR that creates a target site for mir1 and mir206, microRNAs (miRNAs) that are highly expressed in skeletal muscle. This causes translational inhibition of the myostatin gene and hence contributes to the muscular hypertrophy of Texel sheep. Analysis of SNP databases for humans and mice demonstrates that mutations creating or destroying putative miRNA target sites are abundant and might be important effectors of phenotypic variation.

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Figure 1: Mapping and fine-mapping of a QTL influencing muscularity on sheep chromosome 2.
Figure 2: Expression analysis of GDF8 and potentially interacting miRNAs.
Figure 3: Reporter assay testing the interaction between miRNA-GDF8 interaction.

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Acknowledgements

This project was supported by grants from the (i) Walloon Ministry of Agriculture (D31/1036), (ii) the 'GAME' Action de Recherche Concertée from the Communauté Française de Belgique, (iii) the Interuniversity Attraction Pole P5/25 from the Belgian Federal Science Policy Office (R.SSTC.0135), (iv) the European Union 'Callimir' Specific Targeted Research Project (STREP), (v) the University of Liège, (vi) the French Research Agency Genanimal, (vii) Région Auvergne + Départements INRA de Génétique Animale + CEPIA (Caractérisation et élaboration des produits issus de l'agriculture) and (viii) the Région Limousin and Université de Limoges. A.C. benefited from a fellowship of the Département de Génétique Animale, INRA. A.C. and H.T. both benefit from an E.U. Marie-Curie postdoctoral fellowship. C.C. is a 'chercheur qualifié' from the Fonds National de la Recherche Scientifique. We are grateful to P. Leroy and H. Leveziel for their continuous interest and support for this work; to the technical personnel at the Langlade experimental station; to Labogena and France-Upra-Sélection for providing us with DNA samples; to the Centre d'Insémination de Faulx-les-Tombes in Belgium; to C. Fasquelle, V. Marot and V. Dhennin for technical assistance; to J. Vandessompele for advice with real-time PCR and to M. Van Poucke for the primer sequences of the endogenous controls.

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Correspondence to Michel Georges.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

'Within–sire family' QTL analysis demonstrating the TR genotype of the three F1 rams. (PDF 31 kb)

Supplementary Fig. 2

Marker-assisted segregation analysis. (PDF 110 kb)

Supplementary Fig. 3

Comparing the amounts of GDF8 (MSTN) mRNA in skeletal muscle of Texel and wild-type sheep using real-time quantitative RT-PCR. (PDF 65 kb)

Supplementary Fig. 4

SNPs discovered in the ovine GDF8 (MSTN) gene and allelic frequencies in hypermuscled Texels and wild-type controls. (PDF 32 kb)

Supplementary Fig. 5

Sequence context of the polymorphic miRNA-GDF8 interaction in sheep. (PDF 64 kb)

Supplementary Table 1

Effects of the OAR2 QTL on muscularity, fat deposition and body composition significant at the genome-wide 5% level. (PDF 26 kb)

Supplementary Table 2

Genotypes of 42 Texel, 90 controls and four TR rams (three F1, one F2) for the 20 SNPs discovered in the GDF9 (MSTN) gene. (PDF 29 kb)

Supplementary Table 3

Primer sequences. (PDF 19 kb)

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Clop, A., Marcq, F., Takeda, H. et al. A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep. Nat Genet 38, 813–818 (2006). https://doi.org/10.1038/ng1810

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