MKRN3 inhibits the reproductive axis through actions in kisspeptin-expressing neurons.

: The identification of loss-of-function mutations in MKRN3 in patients with central precocious puberty (CPP) in association with the decrease in MKRN3 expression in the medial basal hypothalamus (MBH) of mice prior to the initiation of reproductive maturation suggest that MKRN3 is acting as a 'brake' on GnRH secretion during childhood. In the current study, we investigated the mechanism by which MKRN3 prevents premature manifestation of the pubertal process. We showed that, as in mice, MKRN3 expression is high in the hypothalamus of rats and nonhuman primates early in life, declining as puberty approaches, and is independent of sex steroid hormones. We demonstrated that Mkrn3 is expressed in Kiss1 neurons of the mouse hypothalamic arcuate nucleus (ARC) and that MKRN3 repressed promoter activity of human KISS1 and TAC3, two key stimulators of GnRH secretion. We further showed that MKRN3 has ubiquitinase activity, that this activity is reduced by MKRN3 mutations affecting the RING finger domain, and that these mutations compromised the ability of MKRN3 to repress KISS1 and TAC3 promoter activity. These results indicate that MKRN3 acts to prevent puberty initiation, at least in part, by repressing KISS1 and TAC3 transcription and that this action may involve a MKRN3-directed ubiquitination-mediated mechanism.