A Role for 3-
            <i>O</i>
            -Sulfated Heparan Sulfate in Promoting Human Cytomegalovirus Infection in Human Iris Cells

Baldwin, John R.; Maus, Erika; Zanotti, Brian; Volin, Michael V.; Tandon, Ritesh; Shukla, Deepak; Tiwari, Vaibhav

doi:10.1128/jvi.00109-15

Cited by 20 publications

(39 citation statements)

References 52 publications

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“…Mutant mouse lung endothelial cell lines were from adult mice were mutated for specific sulfotransferase enzymes by a CRISPR-Cas9 based gene editing system 38 45 46 . Since previous studies showed that 3- O -S HS is important for HCMV entry in human iris stromal cells 28 , we analyzed virus replication in Hs3st1 and Hs3st4 (Glucosaminyl 3- O -sulfotransferase 1 and 4, respectively) knockout cell lines as well as the Hs3st1/4 double knockout cell line. At high (5.0) as well as low (0.01) multiplicity of infection (MOI), MCMV growth was significantly reduced in the single Hs3st1 and Hs3st4 knockouts as well as in the double Hs3st1/4 knockouts, indicating that 3- O -sulfation of HS is important for HCMV infection (Fig 9).…”

Section: Resultsmentioning

confidence: 99%

“…This susceptibility to O -desulfated heparins can be transferred to HSV-1 by recombinant transfer of the gene for glycoprotein C (gC-2) from HSV-2 27 . It was recently established that 3- O -S HS are important for HCMV entry in human iris stromal (HIS) cells 28 . The expression of Hs3st in HIS cells promoted HCMV internalization, while pretreatment of HIS cells with heparinase enzyme or treatment with anti-3- O -S HS (G2) peptide significantly reduced HCMV plaques/foci formation.…”

Section: Introductionmentioning

confidence: 99%

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The Degree of Polymerization and Sulfation Patterns in Heparan Sulfate are Critical Determinants of Cytomegalovirus Entry into Host Cells

Hasan

Mitra

Parmar

et al. 2019

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25Herpesviruses attach to host cells by interacting with cell surface heparan sulfate (HS) 26 proteoglycans prior to specific coreceptor engagement which culminates in virus-host 27 membrane fusion and virus entry. Interfering with HS-herpesvirus interactions results in 28 significant reduction in virus infectivity indicating that HS play important roles in initiating 29 virus entry. In this study, we provide convincing evidence that specific sulfations as well 30 as the degree of polymerization (dp) of HS govern human cytomegalovirus (CMV) 31 infection and binding by following line of evidences. First, purified CMV extracellular 32 virions preferentially bound to the sulfated longer chain of HS on a glycoarray compared 33 to unsulfated glycosaminoglycans and shorter chain unsulfated HS. Second, the fraction 34 of glycosaminoglycans (GAG) displaying higher dp and sulfation had a major impact on 35 CMV infectivity and titers. Finally, cell lines knocked out for specific sulfotransferases 36Glucosaminyl 3-O-sulfotransferase (3-O-ST-1 and -4 and double -1/4) produced 37 significantly reduced CMV titers compared to wild-type cells. Similarly, a peptide 38 generated against sulfated-HS significantly reduced virus titers compared to the control 39 peptide. Taken together, the above results highlight the significance of the chain length 40 and sulfation patterns of HS in CMV binding and infectivity. mutant mouse cells and virus binding to glycoarrays. Combined together, the data 50 suggests that virus particles preferentially attach to specifically modified HS and thus 51 the process is amenable to targeting by specifically designed HS mimics. 52 53 54The heparan sulfate (HS) proteoglycans are present on most cell types and function as 55 primary cellular receptor for medically important viruses, including human 56 immunodeficiency virus (HIV), hepatitis-C virus (HCV), human papillomavirus (HPV), 57and Dengue virus (DENV) 1-4 . In addition, virtually all human herpesviruses, with the 58 possible exception of Epstein Barr virus, use HS as an initial co-receptor for entry 5 . The 59 interaction between cell surface HS and virus envelope is the primary event in the 60 complex process of virus entry. However, this binding is not sufficient for viral entry and 61 requires fusion between the viral envelope and cell membrane 6 . 62Herpesviruses including other enveloped viruses enter the host cells using two 63 distinct pathways: 1) A pH-intendent pathway which involves the fusion of the virus 64 envelope with the plasma membrane; and 2) A pH-dependent pathway that involves 65 endocytosis of the virus particle 7 . In cells, where binding of virus to cell surface 66 receptors induces endocytosis, the usual consequence is the acidification of the 67 endosome, which ultimately triggers fusion between the virus envelope and endosomal 68 membrane 5 . Interestingly, HCMV entry follows direct fusion at the cell surface in 69 fibroblasts, while entry into other relevant cell types, such as endothelial cells, follows 70 an endocytic ro...

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Degree of Polymerization and Sulfation Patterns in Heparan Sulfate are Critical Determinants of Cytomegalovirus Entry into Host Cells

Hasan

Mitra

Parmar

et al. 2019

Preprint

Self Cite

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“…While p5R and p5R D reduce HCMV infection in all cell types, there are subtle differences in the percent reduction that potentially reflects differences in HS expression on the cell. It is known that CMV uses a 6-O-sulfated and 3-O-sulfated heparan sulfate for entry (Baldwin et al, 2015; Borst et al, 2013). Our data suggest that p5R D binds to a specific subset of HS, that is different from HS targeted by the previously characterized p5+14 and the HSV-1 entry inhibitor peptide, G2 (Dogra et al, 2015; Tiwari et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…CMVs utilize HSGPs for their initial attachment to the cell and subsequent initiation of infection (Compton et al, 1993). Previous studies have shown that HCMV may preferentially bind to 6-O-sulfated or 3-O-sulfated heparan sulfate moieties during viral entry (Baldwin et al, 2015; Borst et al, 2013). The distribution of HS on host cells in addition to viral preference for specific subtypes of HS, make this structure a potential anti-CMV therapeutic target.…”

Section: Introductionmentioning

confidence: 99%

The D-form of a novel heparan binding peptide decreases cytomegalovirus infection in vivo and in vitro

et al. 2016

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Human cytomegalovirus (HCMV) infection in utero can lead to congenital sensory neural hearing loss and mental retardation. Reactivation or primary infection can increase the morbidity and mortality in immune suppressed transplant recipients and AIDS patients. The current standard of care for HCMV disease is nucleoside analogs, which can be nephrotoxic. In addition resistance to current treatments is becoming increasingly common. In an effort to develop novel CMV treatments, we tested the effectiveness of the D-form of a novel heparan sulfate binding peptide, p5RD, at reducing infection of ganciclovir (GCV) resistant HCMVs in vitro and MCMV in vivo. HCMV infection was reduced by greater than 90% when cells were pretreated with p5RD. Because p5RD acts by a mechanism unrelated to those used by current antivirals, it was effective at reducing GCV resistant HCMVs by 85%. We show that p5RD is resistant to common proteases and serum inactivation, which likely contributed to its ability to significantly reduced infection of peritoneal exudate cells and viral loads in the spleen and the lungs in vivo. The ability of p5RD to reduce HCMV infectivity in vitro including GCV resistant HCMVs and MCMV infection in vivo suggests that this peptide could be a novel anti-CMV therapeutic.

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“…Fondaparinux is a common oligosaccharide used in biophysical studies of GAG interactions with proteins, in which a readily available GAG oligosaccharide of the defined structure is required for the study [17,[18][19][20]. The synthetic pentasaccharide includes a 3-O-sulfated glucosamine, which has been implicated previously in adhesion of some viruses to cell surfaces [21,22]. The experimental characterization and computational modeling of the contact sites between gp120 and fondaparinux will provide a better understanding of how gp120 interacts with fondaparinux and suggest possible methods for blocking the entry of the HIV virus into the host cells.…”

Section: Introductionmentioning

confidence: 99%

Mapping of the Fondaparinux Binding Site of JR-FL gp120 by High Resolution Hydroxyl Radical Protein Footprinting and Computational Docking

Sood

et al. 2017

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The adhesion of HIV gp120 antigen to human cells is modulated in part by interactions with heparan sulfate. The HXB2 strain of gp120 has been shown to interact with heparin primarily through the V3 loop, although other domains including the C-terminal domain were also implicated. However, the JR-FL strain (representative of CCR5-interacting strains that make up newest infections) was shown to have a drastically lowered affinity to heparin due to the loss of several basic residues in the V3 loop, and deletion of the V3 loop in JR-FL gp120 was shown to abrogate some, but not all, heparin binding. Here, we use high resolution hydroxyl radical protein footprinting to measure the changes in protein surface oxidation levels that result from the binding of a model heparin fragment (fondaparinux). Protection in both the V3 loop and the N-terminus of JR-FP gp120 is observed. The well-defined composition of fondaparinux allowed us to perform docking simulations, which showed two clusters of fondaparinux binding: the V3 loop, and a domain consisting of the N-and C-termini. Together, the experimental and theoretical results indicate the heparin/heparan sulfate binding sites on JR-FL gp120 and the efficient interaction of fondaparinux, a widely exploited therapeutic carbohydrate, on gp120.

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A Role for 3- O -Sulfated Heparan Sulfate in Promoting Human Cytomegalovirus Infection in Human Iris Cells

Cited by 20 publications

References 52 publications

The Degree of Polymerization and Sulfation Patterns in Heparan Sulfate are Critical Determinants of Cytomegalovirus Entry into Host Cells

The Degree of Polymerization and Sulfation Patterns in Heparan Sulfate are Critical Determinants of Cytomegalovirus Entry into Host Cells

The D-form of a novel heparan binding peptide decreases cytomegalovirus infection in vivo and in vitro

Mapping of the Fondaparinux Binding Site of JR-FL gp120 by High Resolution Hydroxyl Radical Protein Footprinting and Computational Docking

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