Fujioka Yoshie A, Onuma Asuka, Fujii Wataru, Sugiura Koji, Naito Kunihiko
Laboratory of Applied Genetics, Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
J Reprod Dev. 2018 Jun 22;64(3):253-259. doi: 10.1262/jrd.2018-006. Epub 2018 Mar 23.
Vertebrate oocytes arrested at the first meiotic prophase must proceed to the second meiotic metaphase (MII) before fertilization. This meiotic process requires the precise control of protein degradation. Part of the protein degradation in oocytes is controlled by members of the ubiquitin-conjugating enzyme family, UBE2C and UBE2S, which are known to participate in mono-ubiquitination and poly-ubiquitination, respectively. Although UBE2 enzymes have been well studied in mitosis, their contribution to mammalian oocyte meiosis is relatively unknown and has been studied only in mice. Here, we investigated the contribution of UBE2C and UBE2S to porcine oocyte maturation using an RNA injection method. Overexpression of UBE2S prevented MII arrest of oocytes and led to the formation of a pronucleus (PN) at 48 h of culture. This effect was also observed for prolonged cultures of UBE2C-overexpressing oocytes, suggesting the effectiveness of poly-ubiquitination in the rapid escape from M-phase in porcine oocytes. Although the inhibition of either UBE2C or UBE2S by antisense RNA (asRNA) injection had no effect on oocyte maturation, asRNA-injected oocytes showed inhibited PN formation after parthenogenetic activation. These results indicated that ubiquitination of certain factors by UBE2S and UBE2C plays a role in the escape from MII arrest in porcine oocytes. Further investigations to identify the factors and how mono- and/or poly-ubiquitination contributes to protein degradation could provide a better understanding of UBE2 roles in oocyte maturation.
脊椎动物的卵母细胞在第一次减数分裂前期停滞,必须在受精前进入第二次减数分裂中期(MII)。这个减数分裂过程需要对蛋白质降解进行精确控制。卵母细胞中部分蛋白质降解由泛素结合酶家族成员UBE2C和UBE2S控制,已知它们分别参与单泛素化和多泛素化。尽管UBE2酶在有丝分裂中已得到充分研究,但其对哺乳动物卵母细胞减数分裂的贡献相对未知,且仅在小鼠中进行过研究。在这里,我们使用RNA注射方法研究了UBE2C和UBE2S对猪卵母细胞成熟的贡献。UBE2S的过表达阻止了卵母细胞的MII停滞,并在培养48小时后导致原核(PN)形成。在过表达UBE2C的卵母细胞延长培养中也观察到了这种效应,表明多泛素化在猪卵母细胞从M期快速逃逸中具有有效性。尽管通过反义RNA(asRNA)注射抑制UBE2C或UBE2S对卵母细胞成熟没有影响,但asRNA注射的卵母细胞在孤雌激活后显示出原核形成受到抑制。这些结果表明,UBE2S和UBE2C对某些因子的泛素化在猪卵母细胞从MII停滞中逃逸起作用。进一步研究以确定这些因子以及单泛素化和/或多泛素化如何促进蛋白质降解,可能有助于更好地理解UBE2在卵母细胞成熟中的作用。