Growth impairment resulting from expression of influenza virus M2 protein in Saccharomyces cerevisiae: identification of a novel inhibitor of influenza virus

Kurtz, Stephen E.; Luo, Guangxiang; Hahnenberger, Karen M.; Brooks, Chad S.; Gecha, O.; Ingalls, Kim M.; Numata, Kei-Ichi; Krystal, Mark

doi:10.1128/aac.39.10.2204

Cited by 74 publications

(58 citation statements)

References 36 publications

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“…The majority of novel inhibitors identified to date involve either derivatization of amantadine or another M2-inhibitory compound, BL-1743, which was identified from a yeast-based M2 screen (Duque et al, 2011;Kurtz et al, 1995;Rey-Carrizo et al, 2013, 2014Tu et al, 1996;Wang et al, 2009Wang et al, , 2011aWang et al, , b, 2013aWu et al, 2014). Effective inhibitors of several drug-resistant variants have been identified by this approach, although far fewer hits capable of blocking Asn31 channels have arisen.…”

Section: Iav M2mentioning

confidence: 99%

Viroporins: structure, function and potential as antiviral targets

Scott

Griffin

2015

Journal of General Virology

117

146

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The channel-forming activity of a family of small, hydrophobic integral membrane proteins termed 'viroporins' is essential to the life cycles of an increasingly diverse range of RNA and DNA viruses, generating significant interest in targeting these proteins for antiviral development. Viroporins vary greatly in terms of their atomic structure and can perform multiple functions during the virus life cycle, including those distinct from their role as oligomeric membrane channels. Recent progress has seen an explosion in both the identification and understanding of many such proteins encoded by highly significant pathogens, yet the prototypic M2 proton channel of influenza A virus remains the only example of a viroporin with provenance as an antiviral drug target. This review attempts to summarize our current understanding of the channel-forming functions for key members of this growing family, including recent progress in structural studies and drug discovery research, as well as novel insights into the life cycles of many viruses revealed by a requirement for viroporin activity. Ultimately, given the successes of drugs targeting ion channels in other areas of medicine, unlocking the therapeutic potential of viroporins represents a valuable goal for many of the most significant viral challenges to human and animal health.

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Section: Iav M2mentioning

confidence: 99%

Viroporins: structure, function and potential as antiviral targets

Scott

Griffin

2015

Journal of General Virology

117

146

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“…First, ion channel activity has been observed in three different expression systems, Xenopus oocytes (3,11,12), mammalian cells (5,13), and yeast (14). Second, M 2 channel activity has also been recorded in artificial lipid bilayers from a reconstituted peptide corresponding to the transmembrane domain of the M 2 protein (15) and from purified M 2 protein (16).…”

mentioning

confidence: 99%

Permeation and Activation of the M2 Ion Channel of Influenza A Virus

Mould¹,

Drury²,

Frings³

et al. 2000

Journal of Biological Chemistry

148

166

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The M 2 ion channel protein of influenza A virus is essential for mediating protein-protein dissociation during the virus uncoating process that occurs when the virus is in the acidic environment of the lumen of the secondary endosome. The difficulty of determining the ion selectivity of this minimalistic ion channel is due in part to the fact that the channel activity is so great that it causes local acidification in the expressing cells and a consequent alteration of reversal voltage, V rev . We have confirmed the high proton selectivity of the channel (1.5-2.0 ؋ 10 6 ) in both oocytes and mammalian cells by using four methods as follows: 1) comparison of V rev with proton equilibrium potential; 2) measurement of pH in and V rev while Na The M 2 protein of influenza A virus is thought to function as an ion channel that permits protons to enter virus particles during virion uncoating in endosomes. In addition, in influenza virus-infected cells, the M 2 protein causes the equilibration of pH between the acidic lumen of the trans-Golgi network and the cytoplasm (reviewed in Refs. 1 and 2). The activity of the M 2 ion channel is inhibited by the antiviral drug amantadine (3-5). The mature M 2 protein consists of a 23-residue N-terminal extracellular domain, a single internal hydrophobic domain of 19 residues that acts as a transmembrane domain and forms the pore of the channel, and a 54-residue cytoplasmic tail (6). Chemical cross-linking studies showed the M 2 protein to exist minimally as a homotetramer (7-9). Statistical analysis of the ion channel activity of mixed oligomers also indicated that a homotetramer is the minimal active oligomeric form of the protein (10).Despite the small size of the active M 2 oligomer, several lines of evidence indicate that ion channel activity is intrinsic to the M 2 protein. First, ion channel activity has been observed in three different expression systems, Xenopus oocytes (3, 11, 12), mammalian cells (5, 13), and yeast (14). Second, M 2 channel activity has also been recorded in artificial lipid bilayers from a reconstituted peptide corresponding to the transmembrane domain of the M 2 protein (15) and from purified M 2 protein (16). Thus, due to its structural simplicity, the M 2 ion channel is a potentially useful model for the study of ion channels in general.Although a great deal of evidence indicates H ϩ is the biologically relevant ion for the role of M 2 protein in the life cycle of the influenza virus (1, 3, 17-22), other ions have been shown to be capable of flowing through the channel (12). In addition, the ion selectivity measured for the M 2 channel has been found to differ depending on whether the activity was measured in Xenopus oocytes or mammalian cells. When M 2 protein was expressed in oocytes, V rev was found to differ from the proton equilibrium potential, E Hϩ as [H ϩ ] out was varied (12). On the other hand, when M 2 protein was expressed in MEL cells, V rev was found to agree with E Hϩ (5). In a recent study (23), we found I Hϩ of the M 2 ion channel to...

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“…Therefore, inhibitors of M2 would be expected to reverse this reduction in cell growth rate. Accordingly, BL1743 was able to restore the growth rate of the yeast cells and also inhibited the replication of influenza A/Udorn/72 with an ED 50 of approximately 2 µM in plaque assays (Kurtz et al, 1995). In experiments where M2 was expressed in frog oocytes, BL1743 was found to inhibit the function of this ion channel.…”

Section: Inhibition Of Assembly and Releasementioning

confidence: 94%

Approaches and Strategies for the Treatment of Influenza Virus Infections

Colacino

Staschke

Laver

1999

Antivir Chem Chemother

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Influenza A and B viruses belong to theOrthomyxoviridae family of viruses. These viruses are responsible for severe morbidity and significant excess mortality each year. Infection with influenza viruses usually leads to respiratory involvement and can result in pneumonia and secondary bacterial infections. Vaccine approaches to the prophy-laxis of influenza virus infections have been problematic owing to the ability of these viruses to undergo antigenic shift by exchanging genomic segments or by undergoing antigenic drift, consisting of point mutations in the haemagglutinin (HA) and neuraminidase (NA) genes as a result of an error-prone viral polymerase. Historically, antiviral approaches for the therapy of both influenza A and B viruses have been largely unsuccessful until the elucidation of the X-ray crystallographic structure of the viral NA, which has permitted structure-based drug design of inhibitors of this enzyme. In addition, recent advances in the elucidation of the structure and complex function of influenza HA have resulted in the discovery of a number of diverse compounds that target this viral protein. This review article will focus largely on newer antiviral agents including those that inhibit the influenza virus NA and HA. Other novel approaches that have entered clinical trials or been considered for their clinical utility will be mentioned. Keywords: Influenza virus; haemagglutinin; neuraminidase; amantadine; ribavirin; zanamivir; GS 4104Infection with influenza viruses can result in an infectious disease for which there is no adequate control. Killed influenza vaccines against this virus have been available for nearly 70 years but, on a community-wide basis, the use of these vaccines has not resulted in a significant decrease in the morbidity or mortality of this disease and has not alleviated the severe financial loss which occurs as a result of hospitalizations and lost productivity. Currently, the only compounds approved by the US FDA for the treatment of influenza infections are amantadine and rimantadine, both of which target the M2 ion channel protein of the virus (Pinto et al., 1992). However, these compounds are not effective against influenza B virus, which does not have an M2 ion channel. Also, virus mutants resistant to amantadine or rimantadine can be isolated readily from individuals treated with these drugs Hayden et al., 1989). Recently, it has been suggested that resistance to amantadine can be overcome by mixing amantadine with other amantadine derivatives that interfere with the ion channel function of M2 from amantadine-resistant viruses (Scholtissek et al., 1998). Ribavirin, a broad-spectrum antiviral agent Witkowski et al., 1972;Huffman et al., 1973;Oxford, 1975) has been considered for use against influenza A and B viruses, but the clinical trials of this compound have not been encouraging (Cohen et al., 1976;Togo & McCracken, 1976;Magnussen et al., 1977;Smith et al., 1980;Stein et al., 1987) and it has not been approved for the treatment of influenza virus infection.Hist...

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Growth impairment resulting from expression of influenza virus M2 protein in Saccharomyces cerevisiae: identification of a novel inhibitor of influenza virus

Cited by 74 publications

References 36 publications

Viroporins: structure, function and potential as antiviral targets

Viroporins: structure, function and potential as antiviral targets

Permeation and Activation of the M2 Ion Channel of Influenza A Virus

Approaches and Strategies for the Treatment of Influenza Virus Infections

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