Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Expression Analysis of the Mx Gene and Its Genome Structure in Chickens

  • Yin, C.G. (Life Sciences and Engineering Department, JiNing University) ;
  • Du, L.X. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Li, S.G. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Zhao, G.P. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Zhang, J. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Wei, C.H. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Xu, L.Y. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Liu, T. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences) ;
  • Li, H.B. (National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Science, Chinese Academy of Agricultural Sciences)
  • Received : 2009.09.06
  • Accepted : 2009.12.29
  • Published : 2010.07.01
Download PDF
⟨ Previous Next ⟩

Abstract

Among the known interferon-induced antiviral mechanisms, the Mx pathway is one of the most powerful pathways. The Mx protein has direct antiviral activity and inhibits a wide range of viruses by blocking an early stage of the viral replication cycle. Cloning, characterization, and expression of Mx in vivo and in vitro have been conducted. The chicken Mx gene spans 21 kb and is made up of 14 exons and 13 introns, of which the promoter region was analyzed. The real-time PCR results showed that Mx expression was increased in chicken embryo fibroblasts (CEF) after 12- and 24-h induction with polyI: C. Induction of Mx expression by poly I: C in vivo revealed tissue-specific patterns among the chicken tissues tested. A trace expression of Mx was detected in healthy chicken liver tissues from adult chickens without inducement; the expression levels in the liver, heart, and gizzard were higher than in the muscle and kidney. This is the first report to demonstrate the expression of a glutathione-S-transferase-tagged-Mx fusion protein of 75 KDa, as well as the biological activity tested by SDS-PAGE and western blotting.

Keywords

References

  1. Aebi, M., J. Fah, N. Hurt, C. E. Samuel, D. Thomis, L. Bazzigher, J. Pavlovic, O. Haller and P. Staeheli. 1989. cDNA structures and regulation of two interferon-induced human Mx proteins. Mol. Cell. Biol. 9:5062-5072.
  2. Arnheiter, H., M. Frese, R. Kambadur, E. Meier and O. Haller. 1996. Mx transgenic mice--animal models of health. Curr. Top. Microbiol. Immunol. 206:119-147.
  3. Bazzigher, L., A. Schwarz and P. Staeheli. 1993. No enhanced influenza virus resistance of murine and avian cells expressing cloned duck Mx protein. Virology 195:100-112. https://doi.org/10.1006/viro.1993.1350
  4. Camilla, T. O. Benfield, Jon W. Lyall, Georg Kochs and Laurence S. Tiley. 2008. Asparagine 631 variants of the chicken Mx protein do not inhibit influenza virus replication in primary chicken embryo fibroblasts of in vitro surrogate assays. J. Virol. 7533-7539. https://doi.org/10.1128/JVI.00185-08
  5. Carcamo, J., L. Buckbinder and D. Reinberg. 1991. The initiator directs the assembly of a transcription factor IID-dependent transcription complex. Proc. Natl. Acad. Sci. USA. 88:8052-8056. https://doi.org/10.1073/pnas.88.18.8052
  6. Chang, K. C., G. Goldspink and J. Lida. 1990. Studies in the in vivo expression of the influenza resistance gene Mx by in-situ hybridisation. Arch. Virol. 110:151-164. https://doi.org/10.1007/BF01311285
  7. Collet, B. and C. J. Secombes. 2001. The rainbow trout (Oncorhynchus mykiss) Mx1 promoter. Structural and functional characterization. Eur. J. Biochem. 268:1577-1584. https://doi.org/10.1046/j.1432-1327.2001.02021.x
  8. Di Paolo, C., H. P. Hefti, M. Meli, H. Landis and J. Pavlovic. 1999. Intramolecular backfolding of the carboxyl-terminal end of MxA protein is a prerequisite for its oligomerization. J. Biol. Chem. 274:32071-32078. https://doi.org/10.1074/jbc.274.45.32071
  9. Frese, M., G. Kochs, H. Feldmann, C. Hertkorn and O. Haller. 1996. Inhibition of bunyaviruses, phleboviruses, and hantaviruses by human MxA protein. J. Virol. 70: 915-923.
  10. Garber, E. A., C. I. Rosenblum, H. T. Chute, L. M. Scheidel and H. Chen. 1991. Avian retroviral expression of luciferase. Virology 185:652-660. https://doi.org/10.1016/0042-6822(91)90536-K
  11. Haller, O., H. Arnheiter, J. Lindenmann and I. Gresser. 1980. Host gene influences sensitivity to interferon action selectively for influenza virus. Nature 283:660-662. https://doi.org/10.1038/283660a0
  12. Haller, O., M. Frese, D. Rost, P. A. Nuttall and G. Kochs. 1995. Tick-borne thogoto virus infection in mice is inhibited by the orthomyxovirus resistance gene product Mx1. J. Virol. 69:2596-2601.
  13. Haller, O., G. Kochs and F. Weber. 2007a. Interferon, Mx, and viral countermeasures. Cytokine Growth Factor Rev. 18:425-433. https://doi.org/10.1016/j.cytogfr.2007.06.001
  14. Haller, O., P. Staeheli and G. Kochs. 2007b. Interferon-induced Mx proteins in antiviral host defense. Biochemistry 89:812-818. https://doi.org/10.1016/j.biochi.2007.04.015
  15. Haller, O., S. Stertz and G. Kochs. 2007c. The Mx GTPase family of interferon-induced antiviral proteins. Microbes Infect. 9:1636-1643. https://doi.org/10.1016/j.micinf.2007.09.010
  16. Hug, H., M. Costas, P. Staeheli, M. Aebi and C. Weissmann. 1988. Organization of the murine Mx gene and characterization of its interferon- and virus-inducible promoter. Mol. Cell. Biol. 8:3065-3079.
  17. Jin, H. K., A. Takada, Y. Kon, O. Haller and T. Watanabe. 1999. Identification of the murine Mx2 gene: interferon-induced expression of the Mx2 protein from the feral mouse gene confers resistance to vesicular stomatitis virus. J. Virol. 73:4925-4930.
  18. Kasai, Y., H. Chen and S. J. Flint. 1992. Anatomy of an unusual RNA polymerase II promoter containing a downstream TATA element. Mol. Cell. Biol. 12:2884-2897.
  19. Ko, J. H., H. K. Jin, A. Asano, A. Takada, A. Ninomiya, H. Kida, H. Hokiyama, M. Ohara, M. Tsuzuki, M. Nishibori, M. Mizutani and T. Watanabe. 2002. Polymorphisms and the differential antiviral activity of the chicken Mx gene. Genome Res. 12:595-601. https://doi.org/10.1101/gr.210702.ArticlepublishedonlinebeforeprintinMarch2002
  20. Ko, J. H., A. Takada, T. Mitsuhashi, T. Agui and T. Watanabe. 2004. Native antiviral specificity of chicken Mx protein depends on amino acid variation at position 631. Anim. Genet. 35:119-22. https://doi.org/10.1111/j.1365-2052.2004.01096.x
  21. Lee, J. Y., I. Hirono and T. Aoki. 2000. Cloning and analysis of expression of Mx cDNA in Japanese flounder (Paralichthys olivaceus). Dev. Comp. Immunol. 24:407-415. https://doi.org/10.1016/S0145-305X(99)00060-9
  22. Lee, S. H. and S. M. Vidal. 2002. Functional diversity of Mx proteins: variations on a theme of host resistance to infection. Genome Res. 12:527-530. https://doi.org/10.1101/gr.20102
  23. Li, X. Y., L. J. Qu, Z. C. Hou, J. F. Yao, G. Y. Xu and N. Yang. 2007. Genomic structure and diversity of the chicken Mx gene. Poult. Sci. 86:786-789. https://doi.org/10.1093/ps/86.4.786
  24. Meier, E., G. Kunz, O. Haller and H. Arnheiter. 1990. Activity of rat Mx proteins against a rhabdovirus. J. Virol. 64:6263-6269.
  25. Melen, K., T. Ronni, B. Broni, R. M. Krug, C. H. von Bonsdorff and I. Julkunen. 1992. Interferon-induced Mx proteins form oligomers and contain a putative leucine zipper. J. Biol. Chem. 267:25898-25907.
  26. Pavlovic, J., T. Zurcher, O. Haller and P. Staeheli. 1990. Resistance to influenza virus and vesicular stomatitis virus conferred by expression of human MxA protein. J. Virol. 64:3370-3375.
  27. Ronni, T., S. Matikainen, A. Lehtonen, J. Palvimo, J. Dellis, F. Van Eylen, J. F. Goetschy, M. Horisberger, J. Content and I. Julkunen. 1998. The proximal interferon-stimulated response elements are essential for interferon responsiveness: a promoter analysis of the antiviral MxA gene. J. Interf. Cytok. Res. 18:773-781. https://doi.org/10.1089/jir.1998.18.773
  28. Rothman, J. H., C. K. Raymond, T. Gilbert, P. J. O'Hara and T. H. Stevens. 1990. A putative GTP binding protein homologous to interferon-inducible Mx proteins performs an essential function in yeast protein sorting. Cell 61:1063-1074. https://doi.org/10.1016/0092-8674(90)90070-U
  29. Schumacher, B., D. Bernasconi, U. Schultz and P. Staeheli. 1994. The chicken Mx promoter contains an ISRE motif and confers interferon inducibility to a reporter gene in chick and monkey cells. Virol. 203:144-148. https://doi.org/10.1006/viro.1994.1464
  30. Schumacher, B. and P. Staeheli. 1998. Domains mediating intramolecular folding and oligomerization of MxA GTPase. J. Biol. Chem. 273:28365-28370. https://doi.org/10.1074/jbc.273.43.28365
  31. Staeheli, P., D. Pravtcheva, L. G. Lundin, M. Acklin, F. Ruddle, J. Lindenmann and O. Haller. 1986 Interferon-regulated influenza virus resistance gene Mx is localized on mouse chromosome 16. J. Virol. 58:967-969.
  32. Tazi-Ahnini, R., F. S. di Giovine, A. J. McDonagh, A. G. Messenger, C. Amadou, A. Cox, G. W. Duff and M. J. Cork. 2000. Structure and polymorphism of the human gene for the interferon-induced p78 protein (MX1): evidence of association with alopecia areata in the Down syndrome region. Hum. Genet. 106: 639-645. https://doi.org/10.1007/s004390050037
  33. Zhao, Y., J. E. Nichols, R. Valdez, C. R. Mendelson and E. R. Simpson. 1996. Tumor necrosis factor-alpha stimulates aromatase gene expression in human adipose stromal cells through use of an activating protein-1 binding site upstream of promoter 1.4. Mol. Endocrinol. 10:1350-1357. https://doi.org/10.1210/me.10.11.1350
  34. Zurcher, T., J. Pavlovic and P. Staeheli. 1992a. Mechanism of human MxA protein action: variants with changed antiviral properties. EMBO J. 11:1657-1661.
  35. Zurcher, T., J. Pavlovic and P. Staeheli. 1992b. Nuclear localization of mouse Mx1 protein is necessary for inhibition of influenza virus. J. Virol. 66:5059-5066.

Cited by

  1. Polymorphisms of the Chicken Mx Gene Promoter and Association with Chicken Embryos’ Susceptibility to Virulent Newcastle Disease Virus Challenge vol.2019, pp.None, 2019, https://doi.org/10.1155/2019/1486072