Gong Fanghua, Wang Jing, Li Jie
School of Pharmacy, Wenzhou Medical University, Wenzhou, China; The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.
The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.
Gene. 2017 Nov 30;635:39-45. doi: 10.1016/j.gene.2017.09.018. Epub 2017 Sep 10.
Peroxiredoxin 1 (Prdx1) is a ubiquitously expressed protein in eukaryotic cells, and plays an important role in cell proliferation, differentiation, apoptosis, and redox signaling. Although Prdx1 has been better studied in yeasts and humans, only few Prdx1 genes have been cloned in green algae. The microalga Dunaliella salina (D. salina) is a model for the study of a variety of human cilia-related diseases. In this study, a suppression subtractive hybridization cDNA library of D. salina was constructed, and 6 flagellum-associated genes including D. salina Prdx1 (DsPrdx1) were isolated and identified. A 956bp full-length cDNA of DsPrdx1 was cloned using rapid amplification of cDNA end (RACE). The open reading frame (ORF) of this DNA sequence encodes a polypeptide of 201 amino acids with a predicted molecular weight of 22kDa and a theoretical isoelectric point (pI) of 5.27. Sequence comparison showed that Prdx1 is highly evolutionarily conserved from the unicellular green alga D. salina to human. To our knowledge, this is the first reported full-length sequence of Prdx1 in D. salina. Interestingly, the protein expression of DsPrdx1 was obviously increased during flagellar disassembly in D. salina. Additionally, a yeast two-hybrid assay showed interaction between Prdx1 and RNA, and suggested that DsPrdx1 can protect RNA from degradation by RNase. Taken together, DsPrdx1 not only participates in flagellar disassembly, but also protects RNA from degradation.
过氧化物酶1（Prdx1）是一种在真核细胞中普遍表达的蛋白质，在细胞增殖、分化、凋亡和氧化还原信号传导中发挥重要作用。尽管Prdx1在酵母和人类中已有更深入的研究，但在绿藻中仅克隆到少数Prdx1基因。微藻杜氏盐藻（D. salina）是研究多种人类纤毛相关疾病的模型。在本研究中，构建了杜氏盐藻的抑制性消减杂交cDNA文库，并分离鉴定了包括杜氏盐藻Prdx1（DsPrdx1）在内的6个与鞭毛相关的基因。利用cDNA末端快速扩增（RACE）技术克隆了DsPrdx1的956bp全长cDNA。该DNA序列的开放阅读框（ORF）编码一个由201个氨基酸组成的多肽，预测分子量为22kDa，理论等电点（pI）为5.27。序列比较表明，从单细胞绿藻杜氏盐藻到人类，Prdx1在进化上高度保守。据我们所知，这是首次报道的杜氏盐藻中Prdx1的全长序列。有趣的是，在杜氏盐藻鞭毛解体过程中，DsPrdx1的蛋白表达明显增加。此外，酵母双杂交试验表明Prdx1与RNA之间存在相互作用，并提示DsPrdx1可以保护RNA不被核糖核酸酶降解。综上所述，DsPrdx1不仅参与鞭毛解体，还能保护RNA不被降解。
