Fleurdépine Sophie, Deragon Jean-Marc, Devic Martine, Guilleminot Jocelyne, Bousquet-Antonelli Cécile
CNRS UMR6547 GEEM, Université Blaise Pascal, 63177 Aubière, France and CNRS UMR5096 LGDP, Université de Perpignan Via Domitia, 66860 Perpignan, France.
Nucleic Acids Res. 2007;35(10):3306-21. doi: 10.1093/nar/gkm200. Epub 2007 Apr 25.
Searches in the Arabidopsis thaliana genome using the La motif as query revealed the presence of eight La or La-like proteins. Using structural and phylogenetic criteria, we identified two putative genuine La proteins (At32 and At79) and showed that both are expressed throughout plant development but at different levels and under different regulatory conditions. At32, but not At79, restores Saccharomyces cerevisiae La nuclear functions in non-coding RNAs biogenesis and is able to bind to plant 3'-UUU-OH RNAs. We conclude that these La nuclear functions are conserved in Arabidopsis and supported by At32, which we renamed as AtLa1. Consistently, AtLa1 is predominantly localized to the plant nucleoplasm and was also detected in the nucleolar cavity. The inactivation of AtLa1 in Arabidopsis leads to an embryonic-lethal phenotype with deficient embryos arrested at early globular stage of development. In addition, mutant embryonic cells display a nucleolar hypertrophy suggesting that AtLa1 is required for normal ribosome biogenesis. The identification of two distantly related proteins with all structural characteristics of genuine La proteins suggests that these factors evolved to a certain level of specialization in plants. This unprecedented situation provides a unique opportunity to dissect the very different aspects of this crucial cellular activity.
以La基序作为查询序列在拟南芥基因组中进行搜索,结果显示存在8种La或La样蛋白。利用结构和系统发育标准,我们鉴定出两种假定的真正的La蛋白(At32和At79),并表明这两种蛋白在植物整个发育过程中均有表达,但表达水平和调控条件不同。At32能够恢复酿酒酵母La在非编码RNA生物合成中的核功能,并且能够结合植物3'-UUU-OH RNA,而At79则不能。我们得出结论,这些La核功能在拟南芥中是保守的,并且由At32支持,我们将其重新命名为AtLa1。一致地,AtLa1主要定位于植物核质中,并且在核仁腔中也能检测到。拟南芥中AtLa1的失活导致胚胎致死表型,发育缺陷的胚胎在早期球形期停滞。此外,突变胚胎细胞显示核仁肥大,表明AtLa1是正常核糖体生物合成所必需的。鉴定出两种具有真正La蛋白所有结构特征的远缘相关蛋白,表明这些因子在植物中进化到了一定程度的特化。这种前所未有的情况为剖析这种关键细胞活动的非常不同的方面提供了独特的机会。