Summary
The biological function of a cold-adapted (ca) mutation residing on the PB2 gene of an influenza A/Ann Arbor/6/60 (A/AA/6/60) ca variant virus in the viral replication cycle at 25° C was studied. The viral polypeptide synthesis of A/AA/6/60 ca variant at 25° C was evident approximately 6 hours earlier than the wild type (wt) virus and yielded twice as many products. The quantitative analysis of viral complementary RNA (cRNA), synthesized in the presence of cycloheximide, revealed that A/AA/6/60 ca variant and a single gene reassortant that contains only the PB2 gene of the ca variant with remaining genes of the wt virus produced equal amount of cRNA at 25° and 33° C, which was an amount approximately four fold greater than the wt virus' cRNA synthesized at 25° C. These results strongly suggest that the ca mutation residing on the PB2 gene of A/AA/6/60 ca variant affects the messenger RNA synthesis at 25° C in the primary transcription.
Similar content being viewed by others
References
Barrett, T., Wolstenholme, A. J., Mahy, B. W.: Transcription and replication of influenza virus RNA. Virology98, 211–225 (1979).
Bishop, D. H. L., Obijeski, J. F., Simpson, R. W.: Transcription of the influenza ribonucleic acid genome by a virion polymerase. I. Optimal conditions forin vitro activity of the ribonucleic acid dependent ribonucleic acid polymerase. J. Virol.8, 66–73 (1971).
Braam, J., Ulmanen, I., Krug, R. M.: Molecular model of a eucaryotic transcription complex: Functions and movements of influenza P proteins during capped RNA-primed transcription. Cell.34, 609–618 (1983).
Carrol, A. R., Wagner, R. R.: Role of the membrane (M) protein in endogenous inhibition ofin vitro transcription by vesicular stomatitis virus. J. Virol.29, 134–142 (1979).
Clements, M. L., O'Donnell, S., Levine, M. M., Chanock, R. M., Murphy, B. R.: Dose response of A/Alaska/6/77 (H 3 N 2) cold-adapted reassortant vaccine virus in adalt volunteers: Role of local antibody in resistance to infection with vaccine virus. Infect. Immun.40, 1044–1051 (1983).
Cox, N. J., Maassab, H. F., Kendal, A. P.: Comparative studies of wild-type and cold-mutant (temperature sensitive) influenza viruses: Nonrandon reassortment of genes during preparation of live virus vaccine candidates by recombination at 25° C between recent H 3 N 2 and H 1 N 1 epidemic strains and cold-adapted A/Ann Arbor/6/60. Virology97, 190–194 (1979).
Cox, N. J., Konneck, I., Kendal, A. P., Maassab, H. F.: Genetic and biochemical analysis of the A/Ann Arbor/6/60 cold-adapted mutant. In:Nayak, D. (ed.), Genetic Variation among Influenza Viruses, 639–652. New York: Academic Press 1981.
Cox, N. J., Kendal, A. P.: Genetic stability of A/Ann Arbor/6/60 cold-mutant (temperature-sensitive) live influenza virus genes: Analysis by oligonucleotide mapping of recombinant vaccine strains before and after replication in volunteers. J. Infect. Dis.149, 194–200 (1984).
Glass, S. E., McGeoch, D., Barry, R. D.: Characterization of the mRNA of influenza virus. J. Virol.16, 1435–1443 (1975).
Hay, A. J., Lomniczi, B., Bellamy, A. R., Skehel, J. J.: Transcription of the influenza virus genome. Virology83, 337–355 (1977).
Kendal, A. P., Cox, N. J., Galphin, J. C., Maassab, H. F.: Comparative studies of wild-type and cold-mutant (temperature-sensitive) influenza viruses: Independent segregation of temperature-sensitivity of virus replication from temperature-sensitivity of virion transcriptase activity during recombination of mutant A/Ann Arbor/6/60 with wild-type H 3 N 2 strains. J. gen. Virol.44, 443–456 (1979).
Krug, R. M., Ueda, M., Palese, P.: Temperature-sensitive mutants of influenza WSN virus defective in virus-specific RNA synthesis. J. Virol.16, 790–796 (1975).
Laemmli, U. K.: Cleavage of structural protein during the assembly of the heat of bacteriophage T4. Nature (London)227, 680–685 (1970).
Lamb, A. R., Choppin, P. W.: Synthesis of influenza virus proteins in infected cells: Translation of viral polypeptides, including three P polypeptides from RNA produced by primary transcription. Virology74, 504–519 (1976).
Lamb, R. A., Etkind, P. R., Choppin, P. W.: Evidence for a ninth influenza viral polypeptide. Virology91, 60–78 (1978).
Maassab, H. F.: Adaption and growth characteristics of influenza virus at 25° C. Nature (London)213, 612–614 (1967).
Maassab, H. F.: Biologic and immunologic characteristics of cold-adapted influenza virus. J. Immunol.102, 728–732 (1969).
Maassab, H. F.: Development of variants of influenza virus. In:Barry, R. D., Mahy, B. W. J. (eds.), The Biology of Large RNA Viruses, 542–566. New York: Academic Press 1970.
Maassab, H. F., DeBorde, D. C.: Characterization of influenza A host range mutant. Virology130, 343–350 (1983).
Mark, G. E., Taylor, J. M., Broni, B., Krug, R. M.: Nuclear accumulation of influenza viral RNA transcripts and the effects of cycloheximide, actinomycin D, and α-amamitin. J. Virol.29, 744–752 (1979).
Moritz, A. J., Kunz, C., Hofman, H., Liehi, E., Reev, P., Maassab, H. F.: Studies with a cold-recombinant A/Victoria/3/75 (H3N2) virus. II. Evaluation in adult volunteers. J. Infect. Dis.142, 857–860 (1980).
Mowshowitz, S. L., Ueda, M.: Temperature-sensitive virion transcriptase activity in mutants of WSN influenza virus. Arch. Virol.52, 135–141 (1976).
Odagiri, T., DeBorde, C. D., Maassab, H. F.: Cold-adapted recombinants of influenza A virus in MDCK cells. I. Development and characterization of A/Ann Arbor/6/60 × A/Alaska/6/77 recombinant viruses. Virology119, 82–95 (1982).
Odagiri, T., Smitka, C. W., Maassab, H. F.: Cold-adapted reassortants of influenza A virus in MDCK cells. II. Role of the temperature-sensitive property of cold-adapted in mice. Microbiol. Immunol.27, 203–206 (1983).
Plotch, S. J., Krug, R. M.: Influenza virion transcriptase: Synthesisin vitro of large, polyadenylic acid-containing complementary RNA. J. Virol.21, 24–34 (1977).
Pons, M. W.: Early RNA synthesis in influenza virus-infected cells. Virology76, 855–859 (1977).
Spring, S. B., Maassab, H. F., Kendal, A. P., Murphy, B. R., Chanock, R. M.: Cold-adapted variants of influenza A. II. Comparison of the genetic and biological properties of ts mutations and recombinants of the cold-adapted A/AA/6/60 strain. Arch. Virol.55, 233–246 (1977).
Sugiura, A., Ueda, M., Tobita, K., Enomoto, C.: Further isolation and characterization of temperature-sensitive mutations of influenza virus. Virology65, 363–373 (1975).
Tannock, G. A., Paul, J. A., Barry, R. D.: Relative immunogenicity of the cold-adapted influenza virus A/Ann Arbor/6/60 (A/AA/6/60) recombinants of A/AA/6/60-ca, and parental strains with similar surface antigens. Infect. Immun.43, 457–462 (1984).
Ulmanen, I., Broni, B. A., Krug, R. M.: Role of two of the influenza virus core P proteins in recognizing cap 1 structures (m7GpppNm) on RNAs and in initiating viral RNA transcription. Proc. Natl. Acad. Sci. U.S.A.78, 7355–7359 (1981).
Ulmanen, I., Broni, B., Krug, R. M.: Influenza virus temperature-sensitive cap (m7GpppNm)-dependent endonuclease. J. Virol.45, 27–35 (1983).
Yamane, N., Nakamura, Y., Yuki, M., Odagiri, T., Ishida, N.: Serological evaluation of an influenza A virus cold-adapted reassortant live vaccine, CR-37 (H1N1), in Japanese adult volunteers. J. Hyg. (Camb.)92, 231–242 (1984).
Zvonarjev, A. Y., Ghendon, Y. Z.: Influence of membrane (M) protein on influenza A virus virion transcriptase activityin vitro and its susceptibility to rimantadine. J. Virol.33, 583–586 (1980).
Author information
Authors and Affiliations
Additional information
With 3 Figures
Rights and permissions
About this article
Cite this article
Odagiri, T., Tosaka, A., Ishida, N. et al. Biological characteristics of a cold-adapted influenza A virus mutation residing on a polymerase gene. Archives of Virology 88, 91–104 (1986). https://doi.org/10.1007/BF01310893
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01310893