C-Jun regulates the promoter of small subunit hemocyanin gene of Litopenaeus vannamei. Issue 84 (January 2019)

• Record Type: Journal Article
• Authors: Yang, Peikui; Yao, Defu; Aweya, Jude Juventus; Wang, Fan; Ning, Pei; Li, Shengkang; Ma, Hongyu; Zhang, Yueling
• Is Part Of: Fish & shellfish immunology. Issue 84(2019)
• Page Start: 639
• Page End: 647
• Publication Date: 2019-01
• Subjects: c-Jun -- Alternative promoter -- Regulation -- Hemocyanin -- Litopenaeus vannamei; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2018.10.054
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Hemocyanin (HMC) is a respiratory glycoprotein, which also plays multifunctional non-specific innate immune defense functions in shrimp. However, the transcriptional regulatory mechanisms of the hemocyanin gene expression have not been reported. In the present study, we cloned a 4324 bp fragment of small subunit hemocyanin (HMCs) gene of Litopenaeus vannamei including the 5′-flanking region, from upstream 2475 bp to downstream 1849 bp (exon 1-intron 1-exon 2) by genome walking method. Four deletion constructs were then generated and their promoter activity assessed using the luciferase reporter system. Interestingly, we identified an alternative promoter (+1516/+1849 bp) located in exon 2, which has stronger promoter activity than the full-length or the other constructs. Bioinformatics analyses revealed that the alternative promoter region contains two conserved binding sites of the transcription factor c-Jun. Mutational analysis and electrophoretic mobility shift assay showed that Litopenaeus vannamei c-Jun (Lvc-Jun) binds to the region +1582/+1589 bp and +1831/+1837 bp of the alternative promoter. Furthermore, overexpression of Lvc-Jun significantly increased the alternative promoter activity, while co-transfection with dsRNA-Lvc-Jun significantly reduced the alternative promoter activity of HMCs. Taken together, our present data indicate that the transcription factor Lvc-Jun is essential for the transcriptional regulation of the HMCs gene expression. Highlights: The 5′-flanking region of Litopenaeus vannamei small subunit hemocyanin (HMCs) gene was cloned by genome walking. An alternative promoter was found in exon 2 of HMCs which had stronger promoter activity than other promoter region. The transcription factor Lvc-Jun binds to and transactivates the HMCs alternative promoter.