Activation of influenza A viruses by trypsin treatment

Klenk, Hans‐Dieter; Rott, R.; Orlich, Michaela; Blödorn, Jochen

doi:10.1016/0042-6822(75)90284-6

Cited by 776 publications

(472 citation statements)

References 29 publications

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“…Comparative analysis of the virus proteins by sodium dodecyl sulfate (NaDodSO4)/polyacrylamide gel electrophoresis has shown that biological activation is accompanied by proteolytic cleavage of a precursor envelope glycoprotein, F0, and the production of the glycoprotein F (1, 2), which consists of two disulfide-linked polypeptide chains (3,4). An analogous phenomenon has been observed for the activation of infectivity of influenza virus involving proteolytic cleavage of the precursor polypeptide HAo to two disulfidebonded polypeptide chains, HA1 and HA2 (5,6). Since the activated forms of the fusion protein of Sendai virus (F) and the hemagglutinin of influenza virus (HA) are required for infectivity (and cell Polyacrylamide Gel Electrophoresis.…”

mentioning

confidence: 87%

Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation.

Gething¹,

White²,

Waterfield³

1978

Proc. Natl. Acad. Sci. U.S.A.

241

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The two glycoproteins of Sendai virus, the hemagglutinin-neuraminidase and the fusion protein (F), were separated and purified by affinity chromatography on a Lens culinaris lectin-Sepharose column. F was shown to consist of two disulfide-bonded glycopolypeptide chains, F1 and F2, of molecular weights 51,000 and 11,000, each of which contained 15% carbohydrate by weight. Amino-terminal sequence analysis showed that F2 was blocked and that the hydrophobic sequence NH2-Phe-Phe-Gly-Ala-Val-Ile-Gly-Ile-Ile-Ala-Leu-Gly-Pro-Ala-Thr- was at the amino terminus of F1. This sequence shows identity at six positions with the hydrophobic amino-terminal sequence of the smaller glycopolypeptide chain, HA2, of the hemagglutinin of influenza virus. Both F1 and HA2 are formed by proteolytic cleavage of precursor glycoproteins (Fo, Sendai virus; HAo, influenza virus). Since these cleavages confer infectivity upon both Sendai and influenza viruses and the ability to induce cell-to-cell fusion upon Sendai virus, the hydrophobic NH2-terminal sequences on F1 and HA2 may play a role in fusion of viral and host-cell membranes.

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mentioning

confidence: 87%

Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation.

Gething¹,

White²,

Waterfield³

1978

Proc. Natl. Acad. Sci. U.S.A.

241

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“…The excision of the arginine residue can be accomplished, for example, by a trypsin-like activity followed by a carboxypepticlase B activity. Processing of the precursor hemagglutinin polypeptide with subsequent formation of HA1 and HA2 is required for production of infectious virions, but it is known that this step can also be produced by trypsin (Klenk et al, 1975;Lazarowitz and Choppin, 1975;Klenk, Rott and Orlich, 1977;Bosch et al, 1979). One can easily envisage the possibility for cleavage by trypsin right after the arginine residue to yield "quasi-normal" HA1 and HA2 (and infectious virus) under conditions where the host cellular protease is absent or inactive.…”

Section: The Protein Sequencementioning

confidence: 99%

“…Although both surface antigens change independently, the hemagglutinin (HA) is the antigen against which neutralizing antibodies are directed (Laver and Kilbourne, 1966;Drzenick, Seto and Rott, 1966). The hemagglutinin is responsible for attachment to virus receptors (Hirst, 1942) and is involved in the initial stages of infection (Lazarowtiz and Choppin, 1975;Klenk et al, 1975). Thus the hemagglutinin is considered to be the most important antigen in determining the unique epidemiology of influenza.…”

Section: Introductionmentioning

confidence: 99%

Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA

Jou

Verhoeyen

Devos

et al. 1980

Cell

219

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SummaryThe complete sequence of a hemagglutinin (HA) gene of a recent human influenza A strain, A/Victoria/3/75, is 1768 nucleotides long and contains the information for 567 amino acids. It codes for a signal peptide of 16 amino acids, the HA1 chain of the mature hemagglutinin of 329 amino acids, a connecting region between HA1 and HA2 consisting of a single arginine residue and the HA2 portion of 221 amino acids. The sequence is compared with the hemagglutinin of two members of other subtypes, the human H2 strain A/Jap/305/57 and the avian Havl strain A/FPV/Rostock/34, and with one of the same H3 subtype, A/Memphis/3/72. To align the HA1 chain of different major subtypes several deletions/insertions of single amino acids must be invoked, but two more extensive differences are found at both ends, one leading to an extension of the amino terminal sequence of HA1 and the other (four residues) occurring in the region processed away between HA1 and HA2. Comparison of the HA1 of two H3 strains suggests that drift probably depends on single base mutations, some of which change antigenic determinants. The HA2 region, which apparently is not involved in the immune response, is highly conserved even between different subtypes, and single base substitutions account for all the observed diversity. A hydrophobic segment of 24 residues is present in the same position close to the carboxyl terminus of HA2 in both Victoria and FPV, and presumably functions in implantation into the lipid bilayer. The many conserved features not only in HA2 but also in HA1 suggest a rather rigid architecture for the whole hemagglutinin molecule.

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“…Influenza A virus strain A/PR/8 (H1N1) was kindly donated by Dr. H.-D. Klenk (Institute of Virology, Marburg, Germany) and was propagated and purified as previously outlined in detail (6). Infectivity was assessed by a standard plaque assay as cytopathic effect on confluent cultures of mycoplasma-free Madin Darby canine kidney II cells with a 0.5% agarose overlay (16). Human monocytes (0.5 X 106/ml) were infected or mock-infected by exposure to 2 MOI (multiplicity of infection} A/PR/8-vims (2 plaque-forming units per cell) for 1 h under serum-free conditions (6,7,9).…”

Section: Methodsmentioning

confidence: 99%

Selective induction of monocyte and not neutrophil-attracting chemokines after influenza A virus infection.

Sprenger

Meyer

Kaufmann

et al. 1996

The Journal of Experimental Medicine

120

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SummaryIt is characteristic for virus infections that monocytes/macrophages and lymphocytes infiltrate infected tissue while neutrophils are absent. To understand the mechanisms selectively attracting mononuclear cells in viral diseases, we examined in an influenza A virus model the expression and regulation of chemokines as candidate molecules responsible for the immigration of leukocytes into inflamed tissue. After influenza A virus infection of human monocytes, a rapid expression of the mononuclear cell attracting CC-chemokine genes MIP-1 , MCP-1, and RANTES occurred which was followed by the release ofchemokine proteins. In striking contrast to CC-chemokines, the expression of the prototype neutrophil CXC-chemoattractants IL-8 and GRO-oe was completely suppressed after influenza A infection. The release of other neutrophil chemotactic factors was excluded by nficrochemotaxis assays. These results suggest that the virus-specific induction of mononuclear cell-attracting chemokines accounts for the preferential influx ofmononuclear leukocytes into virus-infected tissue.A hallmark of tissue inflammation is the recruitment, immigration and activation of leukocytes. Gradients of chemotactic factors direct transendothelial migration and movement through the extracellular matrix (1). Most viral diseases are characterized by the development of a specific infiltration consisting predominantly of mononuclear leukocytes while neutrophils are absent as long as no complicating bacterial superinfection occurs. Previous reports show that exposure of monocytes or macrophages to virus results in the release of various proinflammatory cytokines (2-5). Along this line, we demonstrated that an infection of monocytes with influenza A or coxsackie B3 virus induced TNF-o~, IL-1, and IL-6 production (6-9). However, the induction of these proinflammatory cytokines cannot explain the development of characteristic mononuclear leukocyte infiltrations in virally infected tissue.In virus infections, little attention has been focused on the chemokine family and a systematic analysis is still missing. Chemokines are potent chemoattractant cytokines (10, 11) and have to be considered as the main candidate molecules responsible for the selective recruitment of distinct leukocyte populations. Members of the CC-chemokine subfamily, such as MIP-lcl 1 (macrophage inflammatory protein-lc~), MCP-1 (monocyte chemotactic protein-I), and RANTES (regulated upon activation, normal T cell expressed and secreted) preferentially attract monocytes and lymphocytes (12). The CXC-chemokines which contain an ELR-motifpreceding the first cysteine, such as IL-8 (interleukin-8) or GRO-ci (melanoma growth stimulatory activity) are major neutrophil chemoattractants (13).As a first step to study the mechanisms responsible for the generation of a typical virus-induced tissue infiltration consisting predominantly of mononuclear cells, we employed influenza A virus to infect human monocytes. Here we report the selective induction of mononuclear cell attracting chemokine...

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Activation of influenza A viruses by trypsin treatment

Cited by 776 publications

References 29 publications

Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation.

Purification of the fusion protein of Sendai virus: analysis of the NH2-terminal sequence generated during precursor activation.

Complete structure of the hemagglutinin gene from the human influenza A/Victoria/3/75 (H3N2) strain as determined from cloned DNA

Selective induction of monocyte and not neutrophil-attracting chemokines after influenza A virus infection.

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