Hexamethylene Amiloride Blocks E Protein Ion Channels and Inhibits Coronavirus Replication

Overview

Journal Virology

Specialty Microbiology

Date 2006 Jul 4

PMID 16815524

Citations 137

Authors

Lauren Wilson

Peter Gage

Gary Ewart

Affiliations

Soon will be listed here.

Abstract

All coronaviruses encode a small hydrophobic envelope (E) protein, which mediates viral assembly and morphogenesis by an unknown mechanism. We have previously shown that the E protein from Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) forms cation-selective ion channels in planar lipid bilayers (Wilson, L., McKinlay, C., Gage, P., Ewart, G., 2004. SARS coronavirus E protein forms cation-selective ion channels. Virology 330(1), 322-331). We now report that three other E proteins also form cation-selective ion channels. These E proteins were from coronaviruses representative of taxonomic groups 1-3: human coronavirus 229E (HCoV-229E), mouse hepatitis virus (MHV), and infectious bronchitis virus (IBV), respectively. It appears, therefore, that coronavirus E proteins in general, belong to the virus ion channels family. Hexamethylene amiloride (HMA)--an inhibitor of the HIV-1 Vpu virus ion channel--inhibited the HCoV-229E and MHV E protein ion channel conductance in bilayers and also inhibited replication of the parent coronaviruses in cultured cells, as determined by plaque assay. Conversely, HMA had no antiviral effect on a recombinant MHV with the entire coding region of E protein deleted (MHVDeltaE). Taken together, the data provide evidence of a link between inhibition of E protein ion channel activity and the antiviral activity of HMA.

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References

Baudoux P, Carrat C, Besnardeau L, Charley B, Laude H . Coronavirus pseudoparticles formed with recombinant M and E proteins induce alpha interferon synthesis by leukocytes. J Virol. 1998; 72(11):8636-43. PMC: 110275. DOI: 10.1128/JVI.72.11.8636-8643.1998. View

Lu Y, Clavijo P, Galantino M, Shen Z, Liu W, Tam J . Chemically unambiguous peptide immunogen: preparation, orientation and antigenicity of purified peptide conjugated to the multiple antigen peptide system. Mol Immunol. 1991; 28(6):623-30. DOI: 10.1016/0161-5890(91)90131-3. View

Fleming D . Managing influenza: amantadine, rimantadine and beyond. Int J Clin Pract. 2001; 55(3):189-95. View

Madan V, Garcia M, Sanz M, Carrasco L . Viroporin activity of murine hepatitis virus E protein. FEBS Lett. 2005; 579(17):3607-12. PMC: 7094224. DOI: 10.1016/j.febslet.2005.05.046. View

Sakaguchi T, Leser G, Lamb R . The ion channel activity of the influenza virus M2 protein affects transport through the Golgi apparatus. J Cell Biol. 1996; 133(4):733-47. PMC: 2120830. DOI: 10.1083/jcb.133.4.733. View

Shen X, Xue J, Yu C, Luo H, Qin L, Yu X . Small envelope protein E of SARS: cloning, expression, purification, CD determination, and bioinformatics analysis. Acta Pharmacol Sin. 2003; 24(6):505-11. View

Schubert U, Bour S, Ferrer-Montiel A, Montal M, Maldarell F, Strebel K . The two biological activities of human immunodeficiency virus type 1 Vpu protein involve two separable structural domains. J Virol. 1996; 70(2):809-19. PMC: 189883. DOI: 10.1128/JVI.70.2.809-819.1996. View

Ewart G, Sutherland T, Gage P, Cox G . The Vpu protein of human immunodeficiency virus type 1 forms cation-selective ion channels. J Virol. 1996; 70(10):7108-15. PMC: 190763. DOI: 10.1128/JVI.70.10.7108-7115.1996. View

Wilson L, McKinlay C, Gage P, Ewart G . SARS coronavirus E protein forms cation-selective ion channels. Virology. 2004; 330(1):322-31. PMC: 7111769. DOI: 10.1016/j.virol.2004.09.033. View

10.

Fischer F, Stegen C, Masters P, Samsonoff W . Analysis of constructed E gene mutants of mouse hepatitis virus confirms a pivotal role for E protein in coronavirus assembly. J Virol. 1998; 72(10):7885-94. PMC: 110113. DOI: 10.1128/JVI.72.10.7885-7894.1998. View

11.

Pavlovic D, Neville D, Argaud O, Blumberg B, Dwek R, Fischer W . The hepatitis C virus p7 protein forms an ion channel that is inhibited by long-alkyl-chain iminosugar derivatives. Proc Natl Acad Sci U S A. 2003; 100(10):6104-8. PMC: 156333. DOI: 10.1073/pnas.1031527100. View

12.

Maeda J, Repass J, Maeda A, Makino S . Membrane topology of coronavirus E protein. Virology. 2001; 281(2):163-9. PMC: 7130618. DOI: 10.1006/viro.2001.0818. View

13.

Ortego J, Escors D, Laude H, Enjuanes L . Generation of a replication-competent, propagation-deficient virus vector based on the transmissible gastroenteritis coronavirus genome. J Virol. 2002; 76(22):11518-29. PMC: 136772. DOI: 10.1128/jvi.76.22.11518-11529.2002. View

14.

Gonzalez J, Gomez-Puertas P, Cavanagh D, Gorbalenya A, Enjuanes L . A comparative sequence analysis to revise the current taxonomy of the family Coronaviridae. Arch Virol. 2003; 148(11):2207-35. PMC: 7087110. DOI: 10.1007/s00705-003-0162-1. View

15.

Ewart G, Mills K, Cox G, Gage P . Amiloride derivatives block ion channel activity and enhancement of virus-like particle budding caused by HIV-1 protein Vpu. Eur Biophys J. 2002; 31(1):26-35. DOI: 10.1007/s002490100177. View

16.

Fischer W, Sansom M . Viral ion channels: structure and function. Biochim Biophys Acta. 2002; 1561(1):27-45. DOI: 10.1016/s0304-4157(01)00009-0. View

17.

Hout D, Gomez M, Pacyniak E, Gomez L, Fegley B, Mulcahy E . Substitution of the transmembrane domain of Vpu in simian-human immunodeficiency virus (SHIVKU1bMC33) with that of M2 of influenza A results in a virus that is sensitive to inhibitors of the M2 ion channel and is pathogenic for pig-tailed macaques. Virology. 2005; 344(2):541-59. DOI: 10.1016/j.virol.2005.08.022. View

18.

Griffin S, Beales L, Clarke D, Worsfold O, Evans S, Jaeger J . The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine. FEBS Lett. 2003; 535(1-3):34-8. DOI: 10.1016/s0014-5793(02)03851-6. View

19.

Pinto L, Holsinger L, Lamb R . Influenza virus M2 protein has ion channel activity. Cell. 1992; 69(3):517-28. DOI: 10.1016/0092-8674(92)90452-i. View

20.

Hout D, Gomez M, Pacyniak E, Gomez L, Inbody S, Mulcahy E . Scrambling of the amino acids within the transmembrane domain of Vpu results in a simian-human immunodeficiency virus (SHIVTM) that is less pathogenic for pig-tailed macaques. Virology. 2005; 339(1):56-69. DOI: 10.1016/j.virol.2005.04.038. View