• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jvirolPermissionsJournals.ASM.orgJournalJV ArticleJournal InfoAuthorsReviewers
J Virol. Apr 1997; 71(4): 2786–2792.
PMCID: PMC191402

Characterization of a second protein (CM2) encoded by RNA segment 6 of influenza C virus.

Abstract

The biochemical properties of a second protein (CM2) encoded by RNA segment 6 of influenza C virus were investigated. Three forms of CM2 with different electrophoretic mobilities (CM2(0), CM2a, and CM2b) were detected in infected cells by immunoprecipitation with antiserum to the glutathione S-transferase (GST)-CM2 fusion protein. Treatment of infected cells with tunicamycin and digestion of immunoprecipitated proteins with endoglycosidase H or peptide-N-glycosidase F suggested that a mannose-rich oligosaccharide core is added to unglycosylated CM2(0) (Mr, approximately 16,000) to form CM2a (Mr, approximately 18,000) and that the processing of the carbohydrate chain from the high-mannose type to the complex type converts CM2a into CM2b, which is heterogeneous in electrophoretic mobility (Mr, approximately 22,000 to 30,000). Labeling of infected cells with [3H]palmitic acid showed that CM2 is fatty acylated. The fatty acid bond was sensitive to treatment with hydroxylamine and mercaptoethanol, which indicates a labile thioester-type linkage. The CM2 protein was also found to form disulfide-linked dimers and tetramers on sodium dodecyl sulfate-polyacrylamide gels under nonreducing conditions. Trypsin treatment of infected cell surfaces as well as of microsome vesicles from infected cells followed by immunoprecipitation with antiserum to the GST fusion protein containing the 56 C-terminal amino acid residues of CM2 suggested that this C-terminal domain is intracellular and exposed to the cytoplasms of microsomes. Furthermore, evidence that a small amount of CM2 is incorporated into progeny virus particles was obtained by Western blot analysis. These results, altogether, suggest that CM2 is an integral membrane protein with biochemical properties similar to those of influenza A virus M2 and influenza B virus NB proteins.

Full Text

The Full Text of this article is available as a PDF (1.3M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ciampor F, Bayley PM, Nermut MV, Hirst EM, Sugrue RJ, Hay AJ. Evidence that the amantadine-induced, M2-mediated conversion of influenza A virus hemagglutinin to the low pH conformation occurs in an acidic trans Golgi compartment. Virology. 1992 May;188(1):14–24. [PubMed]
  • Elder JH, Alexander S. endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4540–4544. [PMC free article] [PubMed]
  • Fukuda M. Cell surface glycoconjugates as onco-differentiation markers in hematopoietic cells. Biochim Biophys Acta. 1985;780(2):119–150. [PubMed]
  • Holsinger LJ, Lamb RA. Influenza virus M2 integral membrane protein is a homotetramer stabilized by formation of disulfide bonds. Virology. 1991 Jul;183(1):32–43. [PubMed]
  • Hongo S, Sugawara K, Nishimura H, Muraki Y, Kitame F, Nakamura K. Identification of a second protein encoded by influenza C virus RNA segment 6. J Gen Virol. 1994 Dec;75(Pt 12):3503–3510. [PubMed]
  • Lamb RA, Lai CJ. Conservation of the influenza virus membrane protein (M1) amino acid sequence and an open reading frame of RNA segment 7 encoding a second protein (M2) in H1N1 and H3N2 strains. Virology. 1981 Jul 30;112(2):746–751. [PubMed]
  • Lamb RA, Zebedee SL, Richardson CD. Influenza virus M2 protein is an integral membrane protein expressed on the infected-cell surface. Cell. 1985 Mar;40(3):627–633. [PubMed]
  • Mueckler M, Lodish HF. The human glucose transporter can insert posttranslationally into microsomes. Cell. 1986 Feb 28;44(4):629–637. [PubMed]
  • Nakamura K, Homma M, Compans RW. Effect of tunicamycin on the replication of Sendai virus. Virology. 1982 Jun;119(2):474–487. [PubMed]
  • Nishimura H, Sugawara K, Kitame F, Nakamura K, Katsushima N, Moriuchi H, Numazaki Y. A human melanoma cell line highly susceptible to influenza C virus. J Gen Virol. 1989 Jul;70(Pt 7):1653–1661. [PubMed]
  • Pinto LH, Holsinger LJ, Lamb RA. Influenza virus M2 protein has ion channel activity. Cell. 1992 May 1;69(3):517–528. [PubMed]
  • Schmidt MF. Fatty acylation of proteins. Biochim Biophys Acta. 1989 Dec 6;988(3):411–426. [PubMed]
  • Sugawara K, Nakamura K, Homma M. Analyses of structural polypeptides of seven different isolates of influenza C virus. J Gen Virol. 1983 Mar;64(Pt 3):579–587. [PubMed]
  • Sugawara K, Nishimura H, Kitame F, Nakamura K. Antigenic variation among human strains of influenza C virus detected with monoclonal antibodies to gp88 glycoprotein. Virus Res. 1986 Oct;6(1):27–32. [PubMed]
  • Sugawara K, Nishimura H, Hongo S, Kitame F, Nakamura K. Antigenic characterization of the nucleoprotein and matrix protein of influenza C virus with monoclonal antibodies. J Gen Virol. 1991 Jan;72(Pt 1):103–109. [PubMed]
  • Sugawara K, Nishimura H, Hongo S, Muraki Y, Kitame F, Nakamura K. Construction of an antigenic map of the haemagglutinin-esterase protein of influenza C virus. J Gen Virol. 1993 Aug;74(Pt 8):1661–1666. [PubMed]
  • Sugrue RJ, Belshe RB, Hay AJ. Palmitoylation of the influenza A virus M2 protein. Virology. 1990 Nov;179(1):51–56. [PubMed]
  • Sugrue RJ, Hay AJ. Structural characteristics of the M2 protein of influenza A viruses: evidence that it forms a tetrameric channel. Virology. 1991 Feb;180(2):617–624. [PubMed]
  • Takatsuki A, Arima K, Tamura G. Tunicamycin, a new antibiotic. I. Isolation and characterization of tunicamycin. J Antibiot (Tokyo) 1971 Apr;24(4):215–223. [PubMed]
  • Tarentino AL, Maley F. Purification and properties of an endo-beta-N-acetylglucosaminidase from Streptomyces griseus. J Biol Chem. 1974 Feb 10;249(3):811–817. [PubMed]
  • Tarentino AL, Gómez CM, Plummer TH., Jr Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F. Biochemistry. 1985 Aug 13;24(17):4665–4671. [PubMed]
  • Veit M, Herrler G, Schmidt MF, Rott R, Klenk HD. The hemagglutinating glycoproteins of influenza B and C viruses are acylated with different fatty acids. Virology. 1990 Aug;177(2):807–811. [PubMed]
  • Veit M, Klenk HD, Kendal A, Rott R. The M2 protein of influenza A virus is acylated. J Gen Virol. 1991 Jun;72(Pt 6):1461–1465. [PubMed]
  • Williams MA, Lamb RA. Determination of the orientation of an integral membrane protein and sites of glycosylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2-terminal region. Mol Cell Biol. 1986 Dec;6(12):4317–4328. [PMC free article] [PubMed]
  • Williams MA, Lamb RA. Polylactosaminoglycan modification of a small integral membrane glycoprotein, influenza B virus NB. Mol Cell Biol. 1988 Mar;8(3):1186–1196. [PMC free article] [PubMed]
  • Yamashita M, Krystal M, Palese P. Evidence that the matrix protein of influenza C virus is coded for by a spliced mRNA. J Virol. 1988 Sep;62(9):3348–3355. [PMC free article] [PubMed]
  • Yokota M, Nakamura K, Sugawara K, Homma M. The synthesis of polypeptides in influenza C virus-infected cells. Virology. 1983 Oct 15;130(1):105–117. [PubMed]
  • Zebedee SL, Richardson CD, Lamb RA. Characterization of the influenza virus M2 integral membrane protein and expression at the infected-cell surface from cloned cDNA. J Virol. 1985 Nov;56(2):502–511. [PMC free article] [PubMed]
  • Zebedee SL, Lamb RA. Influenza A virus M2 protein: monoclonal antibody restriction of virus growth and detection of M2 in virions. J Virol. 1988 Aug;62(8):2762–2772. [PMC free article] [PubMed]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...