Nicosia Aldo, Costa Salvatore, Tagliavia Marcello, Maggio Teresa, Salamone Monica, Adamo Giorgia, Ragusa Maria Antonietta, Bennici Carmelo, Masullo Tiziana, Mazzola Salvatore, Gianguzza Fabrizio, Cuttitta Angela
Laboratory of Molecular Ecology and Biotechnology, National Research Council-Institute for Marine and Coastal Environment (IAMC-CNR) Detached Unit of Capo Granitola, Torretta Granitola, Trapani, Sicily, Italy.
Dipartimento Scienze e Tecnologie Biologiche Chimiche e Farmaceutiche, University of Palermo, Sicily, Italy.
Cell Stress Chaperones. 2016 May;21(3):535-46. doi: 10.1007/s12192-016-0681-9. Epub 2016 Mar 3.
Gene family encoding cellular nucleic acid binding proteins (CNBP) is well conserved among vertebrates; however, there is limited knowledge in lower organisms. In this study, a CNBP homolog from the red swamp crayfish Procambarus clarkii was characterised. The full-length cDNA of PcCNBP was of 1257 bp with a 5'-untranslated region (UTR) of 63 bp and a 3'-UTR of 331 bp with a poly (A) tail, and an open-reading frame (ORF) of 864 bp encoding a polypeptide of 287 amino acids with the predicted molecular weight of about 33 kDa. The predicted protein possesses 7 tandem repeats of 14 amino acids containing the CCHC zinc finger consensus sequence, two RGG-rich single-stranded RNA-binding domain and a nuclear localization signal, strongly suggesting that PcCNBP was a homolog of vertebrate CNBP. The PcCNBP transcript was constitutively expressed in all tested tissues of unchallenged crayfish, including hepatopancreas, gill, eyestalk, haemocytes, intestine, stomach and cuticle with highest expression in haemocytes, intestine, gills and hepatopancreas. The mRNA expression of PcCNBP in haemocytes was modulated at transcriptional level by different immune challenges, suggesting its involvement in the immune response of P. clarkii during both bacteria and viruses infection.
编码细胞核酸结合蛋白(CNBP)的基因家族在脊椎动物中高度保守;然而,在低等生物中对此了解有限。在本研究中,对来自克氏原螯虾的CNBP同源物进行了表征。PcCNBP的全长cDNA为1257 bp,5'非翻译区(UTR)为63 bp,3'UTR为331 bp并带有poly(A)尾,开放阅读框(ORF)为864 bp,编码一个由287个氨基酸组成的多肽,预测分子量约为33 kDa。预测的蛋白质具有7个包含CCHC锌指共有序列的14个氨基酸的串联重复序列、两个富含RGG的单链RNA结合结构域和一个核定位信号,强烈表明PcCNBP是脊椎动物CNBP的同源物。PcCNBP转录本在未受挑战的小龙虾的所有测试组织中组成性表达,包括肝胰腺、鳃、眼柄、血细胞、肠、胃和表皮,在血细胞、肠、鳃和肝胰腺中表达最高。PcCNBP在血细胞中的mRNA表达在转录水平上受到不同免疫刺激的调节,表明其在克氏原螯虾受到细菌和病毒感染期间的免疫反应中发挥作用。