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Comparative Studies of Wild-Type and ‘Cold-Mutant’ (Temperature Sensitive) Influenza Viruses: Genealogy of the Matrix (M) and Non-structural (NS) Proteins in Recombinant Cold-Adapted H3N2 Viruses

Brian R. Murphy

Susan B. Spring

Hunein F. Maassab

^* WHO Collaborating Center for Influenza, Respiratory Virology Branch, Bureau of Laboratories, Center for Disease Control, Public Health Service, Atlanta, Georgia 30333
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20014
Department of Epidemiology, School of Public Health, University of Michigan, Ann Arbor, Michigan 48104, U.S.A.

The matrix (M) protein of the H2N2 virus A/Ann Arbor/6/60 may be distinguished from M protein of several H3N2 viruses and A/New Jersey/76 (Hsw1N1) by SDS acrylamide gel electrophoresis using a discontinuous buffer system. The smallest RNA (RNA 8) of the A/Ann Arbor/6/60 virus may be distinguished from RNA 8 of several H3N2 viruses by acrylamide gel electrophoresis in 3% or 3.6% gels in the absence of urea, if electrophoresis is done at 30 to 35 °C or 20 °C respectively. Ten clones of conditionally-lethal temperature-sensitive (ts) mutants were studied, which derived their cold-adaption and ts genes from mutant A/Ann Arbor/6/60, and their haemagglutinin from the H3N2 virus A/Scotland/840/74. Each clone was found to derive its M protein from A/Ann Arbor/6/60 mutant, and its RNA 8 from A/Scotland/840/74. The only assignment of genes 7 and 8 consistent with these findings for the recombinants is that in each parent virus (and in the recombinants) gene 7 codes for M protein, and gene 8 for NS protein. Furthermore, it may be concluded from the results that the biologically important ts lesions in the A/Ann Arbor/6/60 mutant parent are not present in the NS gene. In addition to the recombinants of A/Ann Arbor/6/60 and A/Scotland/840/74, five independent ts/cold-adapted recombinants of A/Ann Arbor/6/60 mutant with H3N2 and Hsw1N1 wild-type viruses were examined, and all were found to contain the M protein of the A/Ann Arbor/6/60 mutant parent. This is suggestive that M protein may be at least partially responsible for the cold-adaptation and/or ts properties of the A/Ann Arbor/6/60 mutant and the recombinants.

Received 23 February 1977; accepted 6 May 1977.