Abstract
Ribonucleic acid export 1 (Rae1) is an important nucleoporin that participates in mRNA export during the interphase of higher eukaryotes and regulates the mitotic cell cycle. In this study, small RNA interference technology was used to knockdown Rae1, and immunofluorescence, immunoblotting, and chromosome spreading were used to study the role of Rae1 in mouse oocyte meiotic maturation. We found that Rae1 is a crucial regulator of meiotic maturation of mouse oocytes. After the resumption of meiosis (GVBD), Rae1 was concentrated on the kinetochore structure. The knockdown of Rae1 by a specific siRNA inhibited GVBD progression at 2 h, finally leading to a decreased 14 h polar body extrusion (PBE) rate. However, a comparable 14 h PBE rate was found in the control, and the Rae1 knockdown groups that had already undergone GVBD. Furthermore, we found elevated PBE after 9.5 h in the Rae1 knockdown oocytes. Further analysis revealed that Rae1 depletion significantly decreased the protein level of securin. In addition, we detected weakened kinetochore–microtubule (K-MT) attachments, misaligned chromosomes, and an increased incidence of aneuploidy in the Rae1 knockdown oocytes. Collectively, we propose that Rae1 modulates securin protein levels, which contribute to chromosome alignment, K-MT attachments, and aneuploidy in meiosis.
Highlights
The nuclear pore complex (NPC) is a massive multiprotein complex that is embedded in the nuclear envelope and regulates the nucleocytoplasmic transport of diverse molecules in the eukaryotic interphase
A study in U2OS cells validated that ubiquitin-specific protease 11 (USP11) binds to and modulates the ubiquitination of Ribonucleic acid export 1 (Rae1), thereby regulating the interaction with nuclear mitotic apparatus protein (NuMA), which is critical for normal bipolar spindle formation [7]
We evaluated the effect of Rae1 knockdown on oocyte meiotic maturation
Summary
The nuclear pore complex (NPC) is a massive multiprotein complex that is embedded in the nuclear envelope and regulates the nucleocytoplasmic transport of diverse molecules in the eukaryotic interphase. Rae (homologs in Schizosaccharomyces pombe, Rae1p; Saccharomyces cerevisiae, Gle2p) was initially identified in S. pombe as a nucleoporin involved in poly(A) mRNA export [2,3]. Subsequent studies have shown that in addition to mRNA export, Rae participates in mitotic progression [4]. In HeLa cells, Rae interacts with the nuclear mitotic apparatus protein (NuMA), and this interaction and balance is critically required for normal bipolar spindle assembly [6]. A study in U2OS cells validated that ubiquitin-specific protease 11 (USP11) binds to and modulates the ubiquitination of Rae, thereby regulating the interaction with NuMA, which is critical for normal bipolar spindle formation [7]. Rae interacts with chromosome subunit 1 (SMC1), and disrupting this interaction can result in spindle abnormalities in HeLa cells [8,9]
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