CMAS and ST3GAL4 Play an Important Role in the Adsorption of Influenza Virus by Affecting the Synthesis of Sialic Acid Receptors

Zhao, Yi; Zou, Jiahui; Gao, Qingxia; Xie, Shengsong; Cao, Jiyue; Zhou, Hongbo

doi:10.3390/ijms22116081

Cited by 13 publications

(10 citation statements)

References 38 publications

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“…Binding of both φ-CBM 290 and φ-Siglec7 20 to cytidine monophosphate N- acetylneuraminic acid synthase (CMAS) deficient (CMAS KO ) U937 cells was decreased (Fig. 2b) because CMAS KO cannot produce sialic acid; binding of φ-CBM 290 to CMAS KO was minor (< 5% of binding of φ-CBM 290 to WT) and reminiscent of published observations of binding of sialic acid binding lectins to CMAS KO cells 39 . In all these experiments, we observed no differences in binding between φ-MBP 140 and blank phages (Fig.…”

Section: Resultssupporting

confidence: 78%

Multivalent DNA-encoded lectins on phage enable detecting compositional glycocalyx differences

Lima,

Chenarboo,

Sojitra

et al. 2023

Preprint

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Selective detection of disease-associated changes in the cellular glycocalyx is a foundation of modern targeted therapies. Detecting minor changes in the density and identity of glycans on the cell surface is a technological challenge exacerbated by lack of 1:1 correspondence between cellular DNA/RNA and glycan structures on cell surface. We demonstrate that multivalent displays of up to 300 lectins on DNA-barcoded M13 phage on a liquid lectin array (LiLA), detects subtle differences in composition and density of glycans on cellsex vivoand in immune cells or organs in animals. For example, constructs displaying 73 copies of diCBM40 lectin per 700×5 nm virion (φ-CBM73) exhibit non-linear ON/OFF-like recognition of sialoglycans on the surface of normal and cancer cells. In contrast, a high-valency φ-CBM290 display, or soluble diCBM40, exhibit canonical progressive scaling in binding with increased epitope density; these constructs cannot amplify the subtle differences detected by φ-CBM73. Similarly, multivalent displays of diCBM40 and Siglec-7 detect differences in the glycocalyx between stem-like and non-stem populations in cancer cells that are not detected with soluble lectins. Multivalent display of lectins on M13 scaffold with protected DNA inside the phage offer non-destructive detection of minor differences in glycocalyx in cellsin vitroandin vivonot feasible to currently available technologies.

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Section: Resultssupporting

confidence: 78%

Multivalent DNA-encoded lectins on phage enable detecting compositional glycocalyx differences

Lima,

Chenarboo,

Sojitra

et al. 2023

Preprint

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“…In contrast to previous genome-scale CRISPR screens primarily focused on the isolated human or avian influenza virus in their hosts, our study initiated a genome-scale CRISPR screen in porcine kidney cells to identify the host-dependency factors necessary for EA H1N1 SIV infection 24,28,32 . Shared hits with other studies highlighted the enrichment of host factors involved in SA receptor biosynthesis and related glycosylation pathways, including SLC35A1, CMAS, ST3GAL4, and ALG5, emphasizing the critical role of SA receptor biosynthesis in multiple IAV strains 28,33 . Additionally, unique genes specific to the EA H1N1 SIV strain (HuB/H1N1) and its corresponding host were identified, indicating their specific involvement in EA H1N1 SIV infection.…”

Section: Discussionmentioning

confidence: 80%

GGCX promotes Eurasian avian-like H1N1 swine influenza virus adaption to interspecies receptor binding

Zou,

Jiang,

Xiao

et al. 2024

Preprint

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The Eurasian avian-like (EA) H1N1 swine influenza virus (SIV) possesses the capacity to instigate the next influenza pandemic, owing to its heightened affinity for the human-type α-2,6 sialic acid (SA) receptor. Nevertheless, the molecular mechanisms underlying the switch in receptor binding preferences of EA H1N1 SIV remain elusive. In this study, we conducted a comprehensive genome-wide CRISPR/Cas9 knockout screen utilizing EA H1N1 SIV in porcine kidney cells. Knocking out the enzyme gamma glutamyl carboxylase (GGCX) reduced virus replication in vitro and in vivo by inhibiting the carboxylation modification of viral haemagglutinin (HA) and the adhesion of progeny viruses, ultimately impeding the replication of EA H1N1 SIV. Furthermore, GGCX was revealed to be the determinant of the D225E substitution of EA H1N1 SIV, and GGCX-medicated carboxylation modification of HA 225E contributed to the receptor binding adaption of EA H1N1 SIV to the α-2,6 SA receptor. Taken together, our CRISPR screen has elucidated a novel function of GGCX in the support of EA H1N1 SIV adaption for binding to α-2,6 SA receptor. Consequently, GGCX emerges as a prospective antiviral target against the infection and transmission of EA H1H1 SIV.

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“…Similarly, when the time of NanH treatment was extended up to 4 h, no further decrease in SNA or MAL-II binding was observed compared to a 30 min treatment time (Figure S2b). Residual lectin binding signal may be attributed to trace cell-surface sialic acids that were inaccessible to the sialidase or background SNA binding which has been observed in studies with sialic acid-deficient CMAS knockout cell lines …”

Section: Resultsmentioning

confidence: 96%

“…Residual lectin binding signal may be attributed to trace cell-surface sialic acids that were inaccessible to the sialidase or background SNA binding which has been observed in studies with sialic acid-deficient CMAS knockout cell lines. 43 After enzymatic desialylation using 50 μg/mL NanH, α-2,6linked Neu5DAz was installed by treating desialylated cells with 200 μM CMP-Neu5Daz (1) and 20 μg/mL ST6GAL1 for 2 h. Remodeling desialylated SK-BR-3 and RAJI cells with CMP-Neu5DAz (1) and ST6GAL1 restored SNA binding (Figure 2a,d), whereas SNA binding was higher than control MCF-7, HS-578-T, and U937 cells (Figure 2b,c,e) after remodeling with CMP-Neu5DAz (1) and ST6GAL1. These results demonstrate efficient incorporation of the Neu5DAz probe onto cell-surface glycoconjugates across the various cell lines examined.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Exo-Enzymatic Cell-Surface Glycan Labeling for Capturing Glycan–Protein Interactions through Photo-Cross-Linking

Babulic

Capicciotti

2022

Bioconjugate Chem.

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Tools to interrogate glycoconjugate–protein interactions in the context of living cells are highly attractive for the identification of critically important functional binding partners of glycan-binding proteins. These interactions are challenging to study due to the low affinity and rapid dissociation rates of glycan–protein binding events. The use of photo-cross-linkers to capture glycan–protein interaction complexes has shown great promise for identifying binding partners involved in these interactions. Current methodologies use metabolic oligosaccharide engineering (MOE) to incorporate photo-cross-linking sugars. However, these MOE strategies are not amenable to all cell types and can result in low incorporation and cell-surface display of the photo-cross-linking probe, limiting their utility for studying many types of interactions. We describe here an exo-enzymatic strategy for selectively introducing photo-cross-linking probes into cell-surface glycoconjugates using the recombinant human sialyltransferase ST6GAL1 and a diazirine-linked CMP-Neu5Ac derivative. Probe introduction is highly efficient, amenable to different cell types, and resulted in improved cross-linking when compared to MOE. This exo-enzymatic labeling approach can selectively introduce the photo-cross-linking sugar onto specific glycan epitopes and subclasses by harnessing the specificity of the sialyltransferase employed, underscoring its potential as a tool to interrogate and identify glycoconjugate ligands for diverse glycan-binding proteins.

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CMAS and ST3GAL4 Play an Important Role in the Adsorption of Influenza Virus by Affecting the Synthesis of Sialic Acid Receptors

Cited by 13 publications

References 38 publications

Multivalent DNA-encoded lectins on phage enable detecting compositional glycocalyx differences

Multivalent DNA-encoded lectins on phage enable detecting compositional glycocalyx differences

GGCX promotes Eurasian avian-like H1N1 swine influenza virus adaption to interspecies receptor binding

Exo-Enzymatic Cell-Surface Glycan Labeling for Capturing Glycan–Protein Interactions through Photo-Cross-Linking

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