N- and 6-O-sulfated heparan sulfates mediate internalization of coxsackievirus B3 variant PD into CHO-K1 cells.
Recently, it was demonstrated that the coxsackievirus B3 variant PD (CVB3 PD) is able to infect coxsackievirus-adenovirus receptor (CAR)-lacking cells by using heparan sulfates (HS) as additional receptors (A. E. Zautner, U. Korner, A. Henke, C. Badorff, and M. Schmidtke, J. Virol. 77:10071-10077, 2003). For this study, competition experiments with growth factors binding to known HS sequences as well as with specifically desulfated heparins were performed with Chinese hamster ovary cells (CHO-K1) to determine the structural requirements of HS for interaction with CVB3. Hepatocyte growth factor interacting with HS sequences containing [IdUA-GlcNSO(3)(6OSO(3))](n), but not basic fibroblast growth factor binding to [HexUA-GlcNSO(3)-HexUA-GlcNSO(3)-IdUA(2OSO(3))](n), was shown to compete effectively with CVB3 PD for cell surface HS. Whereas unmodified heparin and 2-O-desulfated heparin strongly inhibited the CVB3 PD-induced cytopathic effect, the antiviral activity was markedly reduced after N-, O- and 6-O-desulfation of heparin. Taken together, these results indicate that 6-O- and N-sulfation of GlcNAc of HS is crucial for HS interaction with CVB3 PD and that the disaccharide [IdUA-GlcNSO(3)(6OSO(3))](n) is involved in viral binding. Results from experiments with various inhibitors of endocytic pathways suggest that HS-mediated virus internalization is pH dependent. Despite the fact that CVB3 PD initiates infection about four times slower by making use of HS as a receptor than by using CAR, the time required for a complete viral life cycle in Chinese hamster ovary cells was independent of the utilized receptor.
Zautner
Andreas E
AE
Institute of Virology and Antiviral Therapy, Medical Centre, Friedrich Schiller University-Jena, Hans-Knoell-Strasse 2, D-07740 Jena, Germany.
Jahn
Birgit
B
Hammerschmidt
Elke
E
Wutzler
Peter
P
Schmidtke
Michaela
M
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D016466:CHO Cells; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D006224:Cricetinae; D003412:Cricetulus; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D004336:Drug Antagonism; D004357:Drug Synergism; D004705:Endocytosis / Q000187:drug effects*; D029822:Enterovirus B, Human / Q000378:metabolism*; D004769:Enterovirus Infections / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006497:Heparan Sulfate / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011991:Receptors, Virus / Q000172:deficiency; D013997:Time Factors
D000925:Anticoagulants; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005346:Fibroblast Growth Factors; D006493:Heparin; D006497:Heparan Sulfate; D017228:Hepatocyte Growth Factor; D011991:Receptors, Virus; C568771:CLMP protein, human