KSHV cell attachment sites revealed by ultra sensitive tyramide signal amplification (TSA) localize to membrane microdomains that are up-regulated on mitotic cells.
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.
['Garrigues|H Jacques|HJ|', 'Rubinchikova|Yelena E|YE|', 'Rose|Timothy M|TM|']
D005456:Fluorescent Dyes D006566:Herpesviridae Infections D019288:Herpesvirus 8, Human D006801:Humans D021962:Membrane Microdomains D018613:Microscopy, Confocal D008938:Mitosis D015854:Up-Regulation D053585:Virus Attachment
Attachment Tyramide signal amplification (TSA) Kaposi's sarcoma-associated herpesvirus (KSHV) Confocal microscopy Mitosis Binding site Human herpesvirus 8 (HHV8)