RT Journal Article SR Electronic T1 A structural variant in the 5’-flanking region of the TWIST2 gene affects melanocyte development in belted cattle JF bioRxiv FD Cold Spring Harbor Laboratory SP 077065 DO 10.1101/077065 A1 Nivedita Awasthi Mishra A1 Cord Drögemüller A1 Vidhya Jagannathan A1 Rémy Bruggmann A1 Julia Beck A1 Ekkehard Schütz A1 Bertram Brenig A1 Steffi Demmel A1 Simon Moser A1 Heidi Signer-Hasler A1 Aldona Pieńkowska-Schelling A1 Claude Schelling A1 Ronald Rongen A1 Stefan Rieder A1 Robert N. Kelsh A1 Nadia Mercader A1 Tosso Leeb YR 2016 UL http://biorxiv.org/content/early/2016/09/23/077065.abstract AB Belted cattle have a circular belt of unpigmented hair and skin around their midsection. The belt is inherited as a monogenic autosomal dominant trait. We mapped the causative variant to a 54 kb segment on bovine chromosome 3. Whole genome sequence data of 2 belted and 130 control cattle yielded only one private genetic variant in the critical interval in the two belted animals. The belt-associated variant was a copy number variant (CNV) involving the quadruplication of a 6 kb non-coding sequence located approximately 16 kb upstream of the TWIST2 gene. Increased copy numbers at this CNV were strongly associated with the belt phenotype in a cohort of 239 cases and 1303 controls (p = 1.3 x 10-278). We hypothesized that the CNV causes aberrant expression of TWIST2 during neural crest development, which might negatively affect melanoblasts. Functional studies showed that ectopic expression of bovine TWIST2 in neural crest in transgenic zebrafish led to a decrease in melanocyte numbers. Our results thus implicate an unsuspected involvement of TWIST2 in regulating pigmentation and reveal a non-coding CNV underlying a captivating Mendelian character.Author Summary Belted cattle, a spontaneous coat color mutant, have been recognized at least 600 years ago. The striking pigmentation pattern probably has arisen in medieval cattle of the Alpine region. The belt still segregates in Brown Swiss cattle and it has become a breed-defining character in the Lakenvelder or Dutch Belted cattle. The belted allele has also been introgressed into Galloways to form the Belted Galloways. We report here the causative genetic variant, a non-coding copy number variant (CNV) upstream of the TWIST2 gene. We hypothesize that the CNV leads to ectopic expression of TWIST2 in the neural crest, which negatively affects melanocyte development. Overexpression of bovine TWIST2 in transgenic zebrafish embryos led to a decrease in melanocyte numbers, which provides functional support for our hypothesis.