title: KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells.
abstract: Cell surface structures initiating attachment of Kaposi's sarcoma-associated herpesvirus (KSHV) were characterized using purified hapten-labeled virions visualized by confocal microscopy with a sensitive fluorescent enhancement using tyramide signal amplification (TSA). KSHV attachment sites were present in specific cellular domains, including actin-based filopodia, lamellipodia, ruffled membranes, microvilli and intercellular junctions. Isolated microdomains were identified on the dorsal surface, which were heterogeneous in size with a variable distribution that depended on cellular confluence and cell cycle stage. KSHV binding domains ranged from scarce on interphase cells to dense and continuous on mitotic cells, and quantitation of bound virus revealed a significant increase on mitotic compared to interphase cells. KSHV also bound to a supranuclear domain that was distinct from microdomains in confluent and interphase cells. These results suggest that rearrangement of the cellular membrane during mitosis induces changes in cell surface receptors implicated in the initial attachment stage of KSHV entry.
authors.affiliation: Seattle Children's Research Institute, 1900 Ninth Ave, 8th Floor, Seattle, WA 98101, USA. Electronic address: Jacques.garrigues@seattlechildrens.org.
authors.forename: H Jacques
authors.initials: HJ
authors.lastname: Garrigues
authors.affiliation: Seattle Children's Research Institute, 1900 Ninth Ave, 8th Floor, Seattle, WA 98101, USA. Electronic address: yrubinchikova@pnwu.edu.
authors.forename: Yelena E
authors.initials: YE
authors.lastname: Rubinchikova
authors.affiliation: Seattle Children's Research Institute, 1900 Ninth Ave, 8th Floor, Seattle, WA 98101, USA; University of Washington, Seattle, WA 98195, USA. Electronic address: timothy.rose@seattlechildrens.org.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Rose
mesh_terms: D005456:Fluorescent Dyes / Q000378:metabolism; D006566:Herpesviridae Infections / Q000503:physiopathology / Q000821:virology*; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D021962:Membrane Microdomains / Q000821:virology*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008938:Mitosis; D015854:Up-Regulation; D053585:Virus Attachment
chemical_list: D005456:Fluorescent Dyes
keywords: Attachment; Binding site; Confocal microscopy; Human herpesvirus 8 (HHV8); Kaposi's sarcoma-associated herpesvirus (KSHV); Mitosis; Tyramide signal amplification (TSA)