Boothby Thomas C, Wolniak Stephen M
Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
BMC Cell Biol. 2011 Oct 13;12:45. doi: 10.1186/1471-2121-12-45.
Many rapidly developing systems rely on the regulated translation of stored transcripts for the formation of new proteins essential for morphogenesis. The microspores of the water fern Marsilea vestita dehydrate as they mature. During this process both mRNA and proteins required for subsequent development are stored within the microspores as they become fully desiccated and enter into senescence. At this point microspores become transcriptionally silent and remain so upon rehydration and for the remainder of spermatogenesis. Transcriptional silencing coupled with the translation of preformed RNA makes the microspore of M. vestita a useful system in which to study post-transcriptional regulation of RNA.
We have characterized the distribution of mRNA as well as several conserved markers of subnuclear bodies within the nuclei of desiccating spores. During this period, nuclear speckles containing RNA were seen to aggregate forming a single large coalescence. We found that aggregated speckles contain several masked mRNA species known to be essential for spermatogenesis. During spermatogenesis masked mRNA and associated speckle proteins were shown to fragment and asymmetrically localize to spermatogenous but not sterile cells. This asymmetric localization was disrupted by RNAi knockdown of the Marsilea homolog of the Exon Junction Complex core component Mago nashi.
A subset of masked mRNA is stored in association with nuclear speckles during the dormant phase of microspore development in M. vestita. The asymmetric distribution of specific mRNAs to spermatogenous but not sterile cells mirrors their translational activities and appears to require the EJC or EJC components. This suggests a novel role for nuclear speckles in the post-transcriptional regulation of transcripts.
许多快速发育的系统依赖于对储存转录本的调控翻译,以形成形态发生所必需的新蛋白质。水蕨类植物中华萍的小孢子在成熟过程中脱水。在此过程中,后续发育所需的mRNA和蛋白质在小孢子完全干燥并进入衰老状态时被储存于其中。此时小孢子转录沉默,在重新水化后以及精子发生的剩余过程中均保持沉默。转录沉默与预先形成的RNA的翻译相结合,使得中华萍的小孢子成为研究RNA转录后调控的有用系统。
我们已对脱水孢子细胞核内mRNA的分布以及核小体的几个保守标记进行了表征。在此期间,可见含有RNA的核斑聚集形成一个单一的大聚集体。我们发现聚集的核斑含有几种已知对精子发生至关重要的隐蔽mRNA种类。在精子发生过程中,隐蔽mRNA和相关的核斑蛋白被证明会片段化并不对称地定位于生精细胞而非不育细胞。这种不对称定位被外显子连接复合体核心组分Mago nashi的中华萍同源物的RNA干扰敲低所破坏。
在中华萍小孢子发育的休眠期,一部分隐蔽mRNA与核斑相关储存。特定mRNA向生精细胞而非不育细胞的不对称分布反映了它们的翻译活性,并且似乎需要外显子连接复合体或其组分。这表明核斑在转录本的转录后调控中具有新作用。