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Adjustment of receptor-binding and neuraminidase substrate specificties in avian–human reassortant influenza viruses

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Abstract

Balanced action of hemagglutinin (HA) and neuraminidase (NA) is an important condition of influenza virus efficient replication, but a role of HA and NA specificities at oligosaccharide level in maintaining such a balance remains poorly studied. Avian virus HA binds exclusively and NA digests efficiently α2–3-sialylated carbohydrate chains, while human virus HA interacts with α2–6 chains and low-active NA cleaves both α2–3- and α2–6-sialosides. Reassortment between viruses leading to appearance of avian virus HA and human virus NA on the virion surface often resulted in decreasing the replicative potential of the formed variants because of disturbance of a functional balance between “alien” HA and NA. A restoration of the reassortant productivity happened due to the appearance of amino acid substitutions in HA and, sometimes, NA. Here, a role of NA and HA oligosaccharide specificities in a restoration of HA–NA functional balance in high-yield passage variants was studied. Postreassortment changes in HA receptor-binding and NA substrate specificities for three reassortant/passage variant virus pairs towards 3′SiaLac, 3′SiaLacNAc, SiaLec, SiaLea, SiaLex, 6′SiaLac, and 6′SiaLacNAc were determined. Selection of the high-yield variants of the human-avian reassortants led either to twofold decrease in the affinity of HA for most α2–3-sialosides and the appearance of affinity for α2–6-sialosides (H3N2 reassortant), or to decreasing the HA affinity for SiaLec and SiaLea (H3N1 reassortant), or to enhancing the ability of NA to discriminate between α2–3/2–6 substrates (H4N1 reassortant). Thus, all postreassortment changes in oligosaccharide specificities of “alien” HA and NA were directed towards their adjustment to each other, but by different manner.

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Abbreviations

biot:

biotin

BODIPY:

4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid

BSA:

bovine serum albumin

HA:

hemagglutinin

HAU:

hemagglutinating unit

Kd :

dissociation constant

Kaff :

affinity constant

NA:

neuraminidase

Neu5Ac:

N-acetylneuraminic acid

OS:

oligosaccharide

PAA:

polyacrylamide

S0 :

initial substrate concentration

V0 :

initial rate of enzymatic hydrolysis

3′SiaLac:

Neu5Acα2–3Galβ1–4Glc

3′SiaLacNAc:

Neu5Acα2–3Galβ1–4GlcNAc

6′SiaLac:

Neu5Acα2–6Galβ1–4Glc

6′SiaLacNAc:

Neu5Acα2–6Galβ1–4GlcNAc

SiaLec :

Neu5Acα2–3Galβ1–3GlcNAc

SiaLea :

Neu5Acα2–3Galβ1–3(Fucα1–4)GlcNAc

SiaLex :

Neu5Acα2–3Galβ1–4(Fucα1–3)GlcNAc

6Su-3′SiaLacNAc:

Neu5Acα2–3Galβ1–4(6-HSO3)GlcNAc

6Su-6′SiaLacNAc:

Neu5Acα2–6Galβ1–4(6-HSO3)GlcNAc

TN buffer:

0.02 M tris-HCl, pH 7.2, with 0.1 M NaCl

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Acknowledgments

The work was supported by grants of Russian Foundation for Basic Research 07-04-00663 and 06-04-48085, and the Russian Academy of Sciences Presidium Program “Molecular and Cell Biology”.

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Authors and Affiliations

  1. Russian Academy of Sciences, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    Yulia Shtyrya, Larisa Mochalova, Galina Voznova & Nicolai Bovin

  2. Russian Academy of Sciences, Engelhardt Institute of Molecular Biology, Vavilova 32, 119991, Moscow, Russia

    Larisa Mochalova

  3. Russian Academy of Medical Sciences, D. I. Ivanovsky Institute of Virology, Gamaleya 16, 123098, Moscow, Russia

    Irina Rudneva, Aleksandr Shilov & Nikolai Kaverin

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  1. Yulia Shtyrya
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  2. Larisa Mochalova
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Correspondence to Larisa Mochalova.

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Shtyrya, Y., Mochalova, L., Voznova, G. et al. Adjustment of receptor-binding and neuraminidase substrate specificties in avian–human reassortant influenza viruses. Glycoconj J 26, 99–109 (2009). https://doi.org/10.1007/s10719-008-9169-x

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