title: Molecular cloning of the pufferfish (Takifugu rubripes) Mx gene and functional characterization of its promoter.
abstract: Mx proteins are members of a family of interferon-inducible genes that are expressed by cells in response to viral infection. They are important determinants of innate immunity against viral infection in vertebrates. We cloned the pufferfish ( Takifugu rubripes) Mx gene and sequenced 80 kb from the Mx locus. The Fugu Mx gene spans 3.4 kb from the transcription start site to the polyadenylation signal, and is made up of 12 exons and 11 introns. The protein sequence encoded by the Fugu Mx gene is 77%, 48%, and 51% identical to that of trout Mx1, chicken Mx, and mouse Mx1 genes, respectively. The Fugu Mx gene is expressed in a variety of tissues, with high expression detected in the heart, gill, kidney, intestine, and brain. Analysis of the 5'-flanking sequence of the gene showed the presence of two interferon-stimulated response elements (ISRE) at positions -51 to 38 and -97 to 85, relative to the transcription start site. The Fugu Mx promoter was inducible by human IFN-beta in the human hepatoma (Huh7) cells and by polyinosinic: polycytidilic acid in the top minnow hepatoma (PLHC-1) cells. Deletion analysis of the promoter showed that both ISREs contributed to inducibility. These results demonstrate that the molecular mechanisms involved in Mx gene regulation are conserved between fish and mammals.
authors.affiliation: Institute of Molecular and Cell Biology, 30 Medical Drive, 117609 Singapore.
authors.forename: Wai Ho
authors.initials: WH
authors.lastname: Yap
authors.forename: Alice
authors.initials: A
authors.lastname: Tay
authors.forename: Sydney
authors.initials: S
authors.lastname: Brenner
authors.forename: Byrappa
authors.initials: B
authors.lastname: Venkatesh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D040342:Genetic Structures; D006160:Guanosine Triphosphate / Q000378:metabolism; D008969:Molecular Sequence Data; D064547:Myxovirus Resistance Proteins; D011401:Promoter Regions, Genetic / Q000502:physiology*; D023721:Tetraodontiformes / Q000235:genetics*
chemical_list: C579070:Mx1 protein, mouse; D064547:Myxovirus Resistance Proteins; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins