title: c-Cbl-mediated selective virus-receptor translocations into lipid rafts regulate productive Kaposi's sarcoma-associated herpesvirus infection in endothelial cells.
abstract: During target cell entry and infection, many enveloped and nonenveloped viruses utilize cell surface receptors that translocate into lipid rafts (LRs). However, the mechanism behind this translocation is not known. Kaposi's sarcoma-associated herpesvirus (KSHV) interacts with the human microvascular dermal endothelial (HMVEC-d) cell surface heparan sulfate (HS), integrins α3β1, αVβ3, and αVβ5, and the amino acid transporter x-CT protein and enters via c-Cbl-bleb-mediated macropinocytosis (Veettil et al., J. Virol. 82:12126-12144, 2008; Veettil et al., PLoS Pathog. 6:e1001238, 2010). Here we have demonstrated that very early during infection (1 min postinfection), c-Cbl induced the selective translocation of KSHV into the LR along with the α3β1, αVβ3, and x-CT receptors but not αVβ5. Activated c-Cbl localized with LRs at the junctional base of macropinocytic blebs. LR-translocated α3β1 and αVβ3 were monoubiquitinated, leading to productive macropinocytic entry, whereas non-LR-associated αVβ5 was polyubiquitinated, leading to clathrin entry that was targeted to lysosomes. c-Cbl knockdown blocked the macropinocytosis and receptor translocation and diverted KSHV to a clathrin-lysosomal noninfectious pathway. Similar results were also seen by LR disruption with MβCD. These studies provide the first evidence that c-Cbl regulates selective KSHV-α3β1, -αVβ3, and -x-CT receptor translocations into the LRs and differential ubiquitination of receptors which are critical determinants of the macropinocytic entry route and productive infection of KSHV. Our studies suggest that interventions targeting c-Cbl and LRs are potential avenues to block KSHV infection of endothelial cells.
authors.affiliation: Department of Microbiology and Immunology, Chicago Medical School, Rosalind Franklin University of Medicine and Science, 3333 Green Bay Road, North Chicago, IL 60064, USA.
authors.forename: Sayan
authors.initials: S
authors.lastname: Chakraborty
authors.forename: Mohanan
authors.initials: M
authors.lastname: ValiyaVeettil
authors.forename: Sathish
authors.initials: S
authors.lastname: Sadagopan
authors.forename: Nitika
authors.initials: N
authors.lastname: Paudel
authors.forename: Bala
authors.initials: B
authors.lastname: Chandran
mesh_terms: D015153:Blotting, Western; D021961:Cell Membrane Structures; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000821:virology*; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D006566:Herpesviridae Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D019288:Herpesvirus 8, Human / Q000472:pathogenicity*; D006801:Humans; D047468:Immunoprecipitation; D039201:Integrin alpha3beta1 / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism*; D055806:Microvessels; D010873:Pinocytosis; D021381:Protein Transport; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D011991:Receptors, Virus / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000378:metabolism; D015398:Signal Transduction; D012867:Skin / Q000166:cytology / Q000378:metabolism / Q000821:virology; D054875:Ubiquitination; D053586:Virus Internalization
chemical_list: D004279:DNA, Viral; D039201:Integrin alpha3beta1; D039302:Integrin alphaVbeta3; D034741:RNA, Small Interfering; D011991:Receptors, Virus; D019030:Receptors, Vitronectin; C064660:integrin alphaVbeta5; D016315:Proto-Oncogene Proteins c-abl