Fig. 3

Schematic overview of bovine CCDC189. Structure of the bovine CCDC189 gene (transcript-ID: ENSBTAT00000045037). The bovine CCDC189 gene encoding the coiled-coil domain containing protein 189 is transcribed from the reverse strand. It consists of nine exons (grey and dark-grey boxes) (a). A mutant allele (red triangle) at 27,138,357 basepairs disrupts a canonical 5′ splice donor variant in the fourth intron. Schematic view of the cDNA sequence of CCDC189 (b). The coiled-coil domain containing protein 189 contains the flagellar C1a complex subunit C1a-32 between amino acids 127 and 221 and a coiled coil domain between amino acids 280 and 308. Red arrows indicate the positions of the primers used for RT-PCR. The mutant allele activates a cryptic splice site in exon 4 that truncates 10 basepairs from exon 4 (red colors) and introduces a frameshift in translation that eventually creates a premature stop codon (c). The mutant protein, if retained, lacks a large fraction of the flagellar C1a complex subunit C1a-32 and the coiled coil domain. Effect of the mutant allele on CCDC189 transcription (d). The RT-PCR products from testis tissue were shorter from two mt/mt (358 bp) than wt/wt (368 bp) bulls. Lane 5 (nc) is the negative PCR control. The length of the bands was defined using the Image Lab version 5.0 software (Bio-Rad Laboratories)