Cuykendall Tawny N, Houston Douglas W
The University of Iowa, Department of Biology, 257 BB, Iowa City, IA 52242, USA.
Development. 2009 Sep;136(18):3057-65. doi: 10.1242/dev.036855. Epub 2009 Aug 12.
Specification of the dorsoventral axis in Xenopus depends on rearrangements of the egg vegetal cortex following fertilization, concomitant with activation of Wnt/beta-catenin signaling. How these processes are tied together is not clear, but RNAs localized to the vegetal cortex during oogenesis are known to be essential. Despite their importance, few vegetally localized RNAs have been examined in detail. In this study, we describe the identification of a novel localized mRNA, trim36, and characterize its function through maternal loss-of-function experiments. We find that trim36 is expressed in the germ plasm and encodes a ubiquitin ligase of the Tripartite motif-containing (Trim) family. Depletion of maternal trim36 using antisense oligonucleotides results in ventralized embryos and reduced organizer gene expression. We show that injection of wnt11 mRNA rescues this effect, suggesting that Trim36 functions upstream of Wnt/beta-catenin activation. We further find that vegetal microtubule polymerization and cortical rotation are disrupted in trim36-depleted embryos, in a manner dependent on Trim36 ubiquitin ligase activity. Additionally, these embryos can be rescued by tipping the eggs 90 degrees relative to the animal-vegetal axis. Taken together, our results suggest a role for Trim36 in controlling the stability of proteins regulating microtubule polymerization during cortical rotation, and subsequently axis formation.
非洲爪蟾背腹轴的特化取决于受精后卵植物皮质的重排,同时伴随着Wnt/β-连环蛋白信号通路的激活。目前尚不清楚这些过程是如何联系在一起的,但已知在卵子发生过程中定位于植物皮质的RNA至关重要。尽管它们很重要,但很少有植物定位的RNA被详细研究过。在这项研究中,我们描述了一种新的定位mRNA——trim36的鉴定,并通过母体功能丧失实验对其功能进行了表征。我们发现trim36在生殖质中表达,并编码一种含三方基序(Trim)家族的泛素连接酶。使用反义寡核苷酸耗尽母体trim36会导致胚胎腹化和组织者基因表达降低。我们表明,注射wnt11 mRNA可挽救这种效应,这表明Trim36在Wnt/β-连环蛋白激活的上游发挥作用。我们进一步发现,在trim36耗尽的胚胎中,植物微管聚合和皮质旋转受到破坏,其方式依赖于Trim36泛素连接酶活性。此外,通过将卵相对于动物-植物轴倾斜90度,可以挽救这些胚胎。综上所述,我们的结果表明Trim36在控制皮质旋转过程中调节微管聚合的蛋白质稳定性以及随后的轴形成中发挥作用。