Molecular cloning, gene expression and methylation status analysis of PIWIL1 in cattle-yaks and the parental generation

The PIWI-like protein 1 (PIWIL1) plays an important role in spermatogenesis and transposon control. In mammals, the absence of the PIWIL1 gene can lead to over-expression of retrotransposons, spermatogenetic arrest, and male infertility. To assess whether the PIWIL1 gene is associated with the male sterility of cattle-yaks, we examined PIWIL1 and LINE-1 (long interspersed element-1) retrotransposon mRNA expression and DNA methylation patterns in the testes of cattle, yaks, and cattle-yaks. Our results indicate that PIWIL1 mRNA expression in the testes of cattle-yaks is significantly lower than that of cattle or yaks. Correlating with decreased gene expression, we also found that the methylation levels of the PIWIL1 gene in cattle-yaks were significantly higher than in cattle or yaks. However, even though LINE-1 element expressed higher mRNA levels in the testes of cattle-yaks than in their parents, DNA methylation patterns were un-altered. Our results suggest that the higher methylation levels of the PIWIL1 gene in cattle-yaks coincides with decreased PIWIL1 mRNA expression, thereby affecting the transposon silencing mechanism and possibly contributing to cattle-yak male sterility.