Identification of virion-associated transcriptional transactivator (VATT) of SGIV ICP46 promoter and their binding site
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RESEARCH
Open Access
Identification of virion-associated transcriptional transactivator (VATT) of SGIV ICP46 promoter and their binding site on promoter Li-Qun Xia1,2,3,4, Jian-Lin Chen1,2,3,4, Hong-Lian Zhang1,2,3,4, Jia Cai1,2,3,4, Sheng Zhou4,5 and Yi-Shan Lu1,2,3,4*
Abstract Background: Iridoviruses are large DNA viruses that cause diseases in fish, amphibians and insects. Singapore grouper iridovirus (SGIV) is isolated from cultured grouper and characterized as a ranavirus. ICP46 is defined to be a core gene of the family Iridoviridae and SGIV ICP46 was demonstrated to be an immediate-early (IE) gene associated with cell growth control and could contribute to virus replication in previous research. Methods: The transcription start site (TSS) and 5′-untranslated region (5′-UTR) of SGIV ICP46 were determined using 5′ RACE. The core promoter elements of ICP46s were analyzed by bioinformatics analysis. The core promoter region and the regulation model of SGIV ICP46 promoter were revealed by the construction of serially deleted promoter plasmids, transfections, drug treat and luciferase reporter assays. The identification of virion-associated transcriptional transactivator (VATT) that interact with SGIV ICP46 promoter and their binding site on promoter were performed by electrophoretic mobility shift assays (EMSA), DNA pull-down assays and mass spectrometry (MS). Results: SGIV ICP46 was found to have short 5′-UTR and a presumptive downstream promoter element (DPE), AGACA, which locates at + 36 to + 39 nt downstream of the TSS. The core promoter region of SGIV ICP46 located from − 22 to + 42 nt relative to the TSS. VATTs were involved in the promoter activation of SGIV ICP46 and further identified to be VP12, VP39, VP57 and MCP. A 10-base DNA sequence “ATGGCTTTCG” between the TSS and presumptive DPE was determined to be the binding site of the VATTs. Conclusion: Our study showed that four VAATs (VP12, VP39, VP57 and MCP) might bind with the SGIV ICP46 promoter and be involved in the promoter activation. Further, the binding site of the VATTs on promoter was a 10-base DNA sequence between the TSS and presumptive DPE. Keywords: Singapore grouper iridovirus (SGIV), ICP46, Promoter, Virion-associated transcriptional transactivator (VATT), Downstream promoter element (DPE)
Background Iridoviruses are large DNA viruses that cause diseases in fish, amphibians, reptiles and insects, and result in significant economic and ecological losses [1, 2]. Similar to other large dsDNA viruses, the gene transcription of iridoviruses can be divided into 3 categories according to * Correspondence: [email protected] 1 Shenzhen Institute of Guangdong Ocean University, Shenzhen City, Guangdong, China 2 College of Fisheries, Guangdong Ocean University, Zhanjiang City, Guangdong, China Full list of author information is available at the end of the article
the sequence: immediate-early (IE), early (E) and late (L), their corresponding protein encoding are very early protein, early protein and late protein [3–5]. As the first step of viral
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