Pereira Roberta V, Gomes Matheus de S, Olmo Roenick P, Souza Daniel M, Jannotti-Passos Liana K, Baba Elio H, Castro-Borges William, Guerra-Sá Renata
Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Morro do Cruzeiro, Ouro Preto, MG, Brazil.
Parasitol Int. 2013 Apr;62(2):199-207. doi: 10.1016/j.parint.2012.12.009. Epub 2013 Jan 8.
NEDD8 is an ubiquitin-like molecule that covalently binds to target proteins through an enzymatic cascade analogous to ubiquitylation. This modifier is known to bind to p53 and p73, as well as all Cullin family proteins, which are essential components of Skp1/Cul-1/F-box protein (SCF)-like Ub ligase complexes. Here, we focused on a genomic analysis of the genes involved in the NEDD8 conjugation pathway in Schistosoma mansoni. The results revealed seven genes related to NEDD8 conjugation that are conserved in Schistosoma japonicum, Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. We performed quantitative RT-PCR (qRT-PCR), which showed differential profiles for Smnedd8, Smapp1, Smuba3, Smube2f, Smdcn1, Smrbx and Smsenp8 throughout the life cycle of S. mansoni. Upregulation was observed in 3-day-old schistosomula and adult worms for all analysed genes. We also analysed the transcription levels of Cullin family members Smp63 and Smp73, and observed upregulation in early schistosomula, while cercariae and adult worms showed expression levels similar to one another. Taken together, these results suggest that the NEDDylation/DeNEDDylation pathway controls important cellular regulators during worm development from cercariae to schistosomula and, finally, to adult.
NEDD8是一种类泛素分子,它通过类似于泛素化的酶促级联反应与靶蛋白共价结合。已知这种修饰剂可与p53和p73以及所有Cullin家族蛋白结合,而这些蛋白是Skp1/Cul-1/F-box蛋白(SCF)样泛素连接酶复合物的重要组成部分。在此,我们重点对曼氏血吸虫中参与NEDD8缀合途径的基因进行了基因组分析。结果揭示了七个与NEDD8缀合相关的基因,这些基因在日本血吸虫、秀丽隐杆线虫、黑腹果蝇和人类中保守。我们进行了定量逆转录聚合酶链反应(qRT-PCR),结果显示在曼氏血吸虫的整个生命周期中,Smnedd8、Smapp1、Smuba3、Smube2f、Smdcn1、Smrbx和Smsenp8具有不同的表达谱。在3日龄的童虫和成虫中,所有分析基因均上调。我们还分析了Cullin家族成员Smp63和Smp73的转录水平,观察到在早期童虫中上调,而尾蚴和成虫的表达水平彼此相似。综上所述,这些结果表明,NEDDylation/去NEDDylation途径在从尾蚴到童虫,最终到成虫的蠕虫发育过程中控制着重要的细胞调节因子。