title: Gilthead seabream (Sparus aurata) Mx gene promoters respond differentially to IPNV and VHSV infections in RTG-2 cells.
abstract: The understanding of virus-host interactions relies on the knowledge of the regulatory mechanisms of the type I interferon (IFN I)-stimulated genes (ISGs). Among ISGs, those coding Mx proteins play a main role due to their direct antiviral activity. The study of these genes in gilthead seabream is interesting, since this species displays high natural resistance to viral diseases, being asymptomatic carrier of infectious pancreatic necrosis virus (IPNV) and viral haemorrhagic septicaemia virus (VHSV). Gilthead seabream has three Mx genes (Mx1, Mx2, and Mx3), encoding proteins with a wide spectrum of antiviral activity. The structure of the three promoters (pMx1, pMx2 and pMx3) has been previously disclosed, and their response to poly I:C in RTG-2 cells characterized. To further analyze these promoters, their response to two viral infections has been evaluated in the present study. For that purpose, RTG-2 cells transiently transfected with the luciferase gene under the control of each promoter were inoculated with either IPNV or VHSV at two different doses. The highest and lowest fold induction values were recorded for pMx2 and pMx3, respectively. The promoter induction was always stronger after VHSV inoculation than in IPNV-inoculated cells. In addition, the higher dose of VHSV tested induced higher response of the three promoters, whereas in IPNV-infected cells the highest induction was recorded after inoculation with the lower viral dose. To further study the response of the Mx2 promoter, RTG-2 cells stably transfected with the luciferase gene under the control of pMx2 were stimulated with poly I:C and subsequently infected with IPNV or VHSV. Interestingly, IPNV infection inhibited the induction caused by poly I:C, suggesting an antagonistic activity of IPNV on Mx2 transcription. In contrast, VHSV infection did not alter the response triggered by poly I:C. These results highlight the specific regulation that controls the activity of each promoter, and support the existence of complex interactions between host cells, specific Mx promoters, and viruses, which are responsible for the final outcome of a viral infection.
authors.affiliation: Universidad de Málaga, Department of Genetics, Spain.
authors.forename: J A
authors.initials: JA
authors.lastname: González-Mariscal
authors.affiliation: Universidad de Málaga, Department of Genetics, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Fernández-Trujillo
authors.affiliation: Universidad de Málaga, Department of Microbiology, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Alonso
authors.affiliation: Universidad de Málaga, Department of Microbiology, Spain.
authors.forename: E
authors.initials: E
authors.lastname: García-Rosado
authors.affiliation: Universidad de Málaga, Department of Genetics, Spain.
authors.forename: M C
authors.initials: MC
authors.lastname: Álvarez
authors.affiliation: Universidad de Málaga, Department of Genetics, Spain. Electronic address: bejar@uma.es.
authors.forename: J
authors.initials: J
authors.lastname: Béjar
mesh_terms: D000818:Animals; D018175:Birnaviridae Infections / Q000503:physiopathology / Q000662:veterinary*; D002460:Cell Line; D005393:Fish Diseases / Q000235:genetics / Q000821:virology*; D018061:Infectious pancreatic necrosis virus; D064547:Myxovirus Resistance Proteins / Q000235:genetics*; D029141:Novirhabdovirus; D011401:Promoter Regions, Genetic; D018353:Rhabdoviridae Infections / Q000662:veterinary*; D021541:Sea Bream; D014162:Transfection / Q000662:veterinary
chemical_list: D064547:Myxovirus Resistance Proteins
keywords: Antagonism; Gilthead seabream; IPNV; Mx protein; Promoter; VHSV