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J Virol, March 1998, p. 2449-2455, Vol. 72, No. 3
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The M2 Ectodomain Is Important for Its Incorporation into Influenza A Virions

Eun K. Park,1 Maria R. Castrucci,2 Allen Portner,1,3 and Yoshihiro Kawaoka1,3,*

Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 381051; Department of Virology, Instituto Superiore di Sanita, 00161 Rome, Italy2; and Department of Pathology, University of Tennessee--- Memphis, Memphis, Tennessee 381633

Received 22 August 1997/Accepted 12 November 1997

M2 is an integral protein of influenza A virus that functions as an ion channel. The ratio of M2 to HA in influenza A virions differs from that found on the cell surface, suggesting selective incorporation of M2 and HA into influenza virions. To examine the sequences that are important for M2 incorporation into virions, we used an incorporation assay that involves expressing M2 from a plasmid, transfecting the plasmid into recipient cells, and then infecting those cells with influenza virus. To test the importance of the different regions of the protein (extracellular, transmembrane, and cytoplasmic) in determining M2 incorporation, we created chimeric mutants of M2 and Sendai virus F proteins, exchanging corresponding extracellular, transmembrane, and cytoplasmic domains. Of the six possible chimeric mutants, only three were expressed on the cell surface. Of these three chimeric proteins, only one mutant (with the extracellular domain from M2 and the rest from F) was incorporated into influenza virions. These results suggest that the extracellular domain of M2 is important for its incorporation into virions.

* Corresponding author. Present address: Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin---Madison, 2015 Linden Dr. West, Madison, WI 53706. Phone: (608) 265-4925. Fax: (608) 265-5622. E-mail: kawaokay{at}svm.vetmed.wisc.edu.




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