Abstract
We developed a novel drug screening system for anti-influenza A virus by targeting the M2 proton channel. In the SPP (Single Protein Production) system, E. coli cell growth occurs only in the presence of effective M2 channel inhibitors, and thus simple measurement of cell growth was used as readouts for drug screening. Two potential inhibitors for M2 (V27A) mutant were verified using this method, which inhibit both the mutant and wild-type M2 channels.
Highlights
Influenza A virus, which causes flu, is known as one of the most prevalent pathogenic viruses
In the single protein production (SPP) system, the synthesis of all cellular proteins is inhibited by the induction of MazF, an ACA-specific mRNA interferase from Escherichia coli, except for the synthesis of a designated protein, of which mRNA is designed to have no ACA sequences [7]
An ACA-less gene coding for M2 protein was synthesized and cloned into pColdII(sp-4) [7]
Summary
Influenza A virus, which causes flu, is known as one of the most prevalent pathogenic viruses. Compounds 10, 34, 35, 282 and 293 were identified to be potential inhibitors of influenza A virus infection. AcGFP fails to be as a reporter since it is not fluorescent in the AcGFP-M2 fusion protein, we found in the present manuscript that cell growth is resumed as measured by the increase of cell density using O.D.600, when AcGFP-M2 fusion protein is expressed in the presence of inhibitors for the M2 channel activity (Figure 1D).
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