| Summary: | The 3′ untranslated region has an important role in gene regulation through microRNAs, and it has been estimated that microRNAs regulate up to 50% of coding genes in mammals. With the aim of allelic variant identification of 3′ untranslated region microRNA seed sites, the 3′ untranslated region was searched for seed sites of four temperament-associated genes (<i>CACNG4</i>, <i>EXOC4</i>, <i>NRXN3</i>, and <i>SLC9A4</i>). The microRNA seed sites were predicted in the four genes, and the <i>CACNG4</i> gene had the greatest number with 12 predictions. To search for variants affecting the predicted microRNA seed sites, the four 3′ untranslated regions were re-sequenced in a Brahman cattle population. Eleven single nucleotide polymorphisms were identified in the <i>CACNG4</i>, and eleven in the <i>SLC9A4</i>. Rs522648682:T>G of the <i>CACNG4</i> gene was located at the predicted seed site for <i>bta-miR-191</i>. Rs522648682:T>G evidenced an association with both exit velocity (<i>p</i> = 0.0054) and temperament score (<i>p</i> = 0.0097). The genotype TT had a lower mean exit velocity (2.93 ± 0.4 m/s) compared with the TG and GG genotypes (3.91 ± 0.46 m/s and 3.67 ± 0.46 m/s, respectively). The allele associated with the temperamental phenotype antagonizes the seed site, disrupting the <i>bta-miR-191</i> recognition. The G allele of <i>CACNG4</i>-rs522648682 has the potential to influence bovine temperament through a mechanism associated with unspecific recognition of <i>bta-miR-191</i>.
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