Structural and functional characterization of neuraminidase-like molecule N10 derived from bat influenza A virus

Li, Qing; Sun, Xiaoman; Li, Zhixin; Liu, Yue; Vavricka, Christopher J.; Qi, Jianxun; Gao, George F.

doi:10.1073/pnas.1211037109

Cited by 108 publications

(94 citation statements)

References 45 publications

(55 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, the recently reported N10 NAL structures, including GU10-060 NAL and GU09-164 NAL, from bat H17N10 viruses revealed a highly diverged putative active site that is wider than other NAs and in which most of the residues required for NA activity are substituted. These structural features are consistent with the recombinant N10 proteins exhibiting no, or extremely low, NA activity (3,4). Thus, these observations have raised further interest in the structure and function of the HA and NA proteins of bat H17N10 viruses.…”

Section: Discussionsupporting

confidence: 59%

Hemagglutinin homologue from H17N10 bat influenza virus exhibits divergent receptor-binding and pH-dependent fusion activities

Zhu¹,

Yu²,

McBride³

et al. 2013

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

144

110

View full text Add to dashboard Cite

Bat influenza virus H17N10 represents a distinct lineage of influenza A viruses with gene segments coding for proteins that are homologs of the surface antigens, hemagglutinin (HA) and neuraminidase (NA). Our recent study of the N10 NA homolog revealed an NA-like structure, but with a highly divergent putative active site exhibiting little or no NA activity, and provided strong motivation for performing equivalent structural and functional analyses of the H17 HA protein. The overall structure of the H17 HA homolog from A/little yellow-shouldered bat/Guatemala/060/2010 at 3.18 Å resolution is very similar to other influenza HAs, with a putative receptor-binding site containing some conserved aromatic residues that form the base of the sialic acid binding site. However, the rest of the H17 receptor-binding site differs substantially from the other HA subtypes, including substitution of other conserved residues associated with receptor binding. Significantly, electrostatic potential analyses reveal that this putative receptor-binding site is highly acidic, making it unfavorable to bind any negatively charged sialylated receptors, consistent with the recombinant H17 protein exhibiting no detectable binding to sialylated glycans. Furthermore, the fusion mechanism is also distinct; trypsin digestion with recombinant H17 protein, when exposed to pH 4.0, did not degrade the HA1 and HA2, in contrast to other HAs. These distinct structural features and functional differences suggest that the H17 HA behaves very differently compared with other influenza HAs.crystal structure | evolution | infection | protease susceptibility | viral entry I nfluenza is an infectious disease of birds and mammals caused by influenza virus, an RNA virus of the Orthomyxoviridae family. Three types of influenza virus, A, B, and C, are known. Influenza A virus infects a wide range of avian and mammalian species and is the major cause of annual human epidemics and occasional pandemics. Based on the antigenic properties of the two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA), type A viruses can be classified into multiple subtypes. With the recent discovery of the H17N10 subtype of bat influenza A viruses (1), 17 subtypes of HA (H1-17) and 10 subtypes of NA (N1-10) have been found to circulate in avian and/or mammalian hosts. The H17N10 virus expands the species that can be infected by influenza virus to bats, which account for approximately onefourth of all mammalian species and have been shown to be a reservoir for multiple emerging viruses (2).In influenza virus infection, the HA is responsible for binding of virus to sialic acid-containing receptors on host cell surface glycoproteins and glycolipids, internalization of the virus, and subsequently membrane fusion within the endosome of the infected cell. Following virus replication, the receptor-cleavage enzyme, NA, removes sialic acid from glycans on target cell surfaces as well as from newly formed budding virions so that the progeny viruses can be released and infect other...

show abstract

Section: Discussionsupporting

confidence: 59%

Hemagglutinin homologue from H17N10 bat influenza virus exhibits divergent receptor-binding and pH-dependent fusion activities

Zhu¹,

Yu²,

McBride³

et al. 2013

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

144

110

View full text Add to dashboard Cite

show abstract

“…IAVs are classified based on the subtypes of both HA and NA. So far, 18 HA (H1 to H18) and 11 NA (N1 to N11) subtypes have been identified, almost all of which have been found in aquatic birds (3)(4)(5)(6). In humans, IAVs cause seasonal epidemics and occasional pandemics.…”

mentioning

confidence: 99%

Identification of Residues That Affect Oligomerization and/or Enzymatic Activity of Influenza Virus H5N1 Neuraminidase Proteins

Dai

Guo

Dortmans

et al. 2016

J Virol

View full text Add to dashboard Cite

Influenza A virus (IAV) attachment to and release from sialoside receptors is determined by the balance between hemagglutinin (HA) and neuraminidase (NA). The molecular determinants that mediate the specificity and activity of NA are still poorly understood. In this study, we aimed to design the optimal recombinant soluble NA protein to identify residues that affect NA enzymatic activity. To this end, recombinant soluble versions of four different NA proteins from H5N1 viruses were compared with their full-length counterparts. The soluble NA ectodomains were fused to three commonly used tetramerization domains. Our results indicate that the particular oligomerization domain used does not affect the K m value but may affect the specific enzymatic activity. This particularly holds true when the stalk domain is included and for NA ectodomains that display a low intrinsic ability to oligomerize. NA ectodomains extended with a Tetrabrachion domain, which forms a nearly parallel four-helix bundle, better mimicked the enzymatic properties of full-length proteins than when other coiled-coil tetramerization domains were used, which probably distort the stalk domain. Comparison of different NA proteins and mutagenic analysis of recombinant soluble versions thereof resulted in the identification of several residues that affected oligomerization of the NA head domain (position 95) and therefore the specific activity or sialic acid binding affinity (K m value; positions 252 and 347). This study demonstrates the potential of using recombinant soluble NA proteins to reveal determinants of NA assembly and enzymatic activity. IMPORTANCEThe IAV HA and NA glycoproteins are important determinants of host tropism and pathogenicity. However, NA is relatively understudied compared to HA. Analysis of soluble versions of these glycoproteins is an attractive way to study their activities, as they are easily purified from cell culture media and applied in downstream assays. In the present study, we analyzed the enzymatic activity of different NA ectodomains with three commonly used tetramerization domains and compared them with fulllength NA proteins. By performing a mutagenic analysis, we identified several residues that affected NA assembly, activity, and/or substrate binding. In addition, our results indicate that the design of the recombinant soluble NA protein, including the particular tetramerization domain, is an important determinant for maintaining the enzymatic properties within the head domain. NA ectodomains extended with a Tetrabrachion domain better mimicked the full-length proteins than when the other tetramerization domains were used. V irus particles contain dedicated proteins that recognize cell surface molecules. While some viruses evolved to bind specific protein receptors, others bind carbohydrate moieties, such as sialic acids (SIAs), that are omnipresent on the cell surface as well as in the mucus. Several of the latter viruses, including influenza A virus (IAV), carry receptor-destroying activities in addit...

show abstract

“…This suggests that H17N10 PAn is most likely able to generate the short primers needed for the initiation of viral replication via its endonuclease activity. However, for the glyco- proteins from H17N10, HA and NA, no canonical function could be demonstrated, despite their structural similarities to previously characterized HA and NA molecules (9)(10)(11)(12). Therefore, it was interesting to determine whether PAn from H17N10 would indeed have a function similar to that of categorized influenza virus PAn molecules.…”

Section: Discussionmentioning

confidence: 99%

“…Crystal structures for the surface glycoproteins hemagglutinin (HA) (7,9,10) and neuraminidase (NA) (7,11,12) were quickly resolved for these viruses, and, surprisingly, both bat-derived influenza virus proteins were shown to be functionally and structurally distinct from all known HA and NA proteins. Their HA protein did not bind to any sialic acid-or other sugar-containing moiety tested, and their NA did not hydrolyze any substrate containing sialic acids.…”

mentioning

confidence: 99%

The N-Terminal Domain of PA from Bat-Derived Influenza-Like Virus H17N10 Has Endonuclease Activity

Tefsen

Zhu

et al. 2014

J Virol

Self Cite

View full text Add to dashboard Cite

e Influenza imposes a great burden on society, not only in its seasonal appearance that affects both humans and domesticated animals but also through the constant threat of potential pandemics. Migratory birds are considered to be the reservoir hosts for influenza viruses, but other animals must also be considered. The recently identified influenza-like virus genome, from H17N10 in bats, was shown to be markedly different from genomes of other known influenza viruses, as both its surface glycoproteins hemagglutinin (HA) and neuraminidase (NA) do not have canonical functions. However, no studies on other individual proteins from this particular virus have been reported until now. Here, we describe the structure of the N-terminal domain of PA from H17N10 influenza-like virus at 2.7-Å resolution and show that it has a fold similar to those of homologous PA domains present in more familiar influenza A virus strains. Moreover, we demonstrate that it possesses endonuclease activity and that the histidine residue in the active site is essential for this activity. Although this particular influenza virus subtype is probably not infectious for humans (even its virus state has not been confirmed in bats, as only the genome has been sequenced), reassortment of canonical influenza viruses with certain segments from H17N10 cannot be ruled out at this stage. Therefore, further studies are urgently needed for the sake of influenza prevention and control.

show abstract

Structural and functional characterization of neuraminidase-like molecule N10 derived from bat influenza A virus

Cited by 108 publications

References 45 publications

Hemagglutinin homologue from H17N10 bat influenza virus exhibits divergent receptor-binding and pH-dependent fusion activities

Hemagglutinin homologue from H17N10 bat influenza virus exhibits divergent receptor-binding and pH-dependent fusion activities

Identification of Residues That Affect Oligomerization and/or Enzymatic Activity of Influenza Virus H5N1 Neuraminidase Proteins

The N-Terminal Domain of PA from Bat-Derived Influenza-Like Virus H17N10 Has Endonuclease Activity

Contact Info

Product

Resources

About