Fluorescent tagging and cellular distribution of the Kaposi's sarcoma-associated herpesvirus ORF45 tegument protein

Shir Bergson, Inna Kalt, Inbal Itzhak, Kevin F. Brulois, Jae U. Jung, Ronit Sarid

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Kaposi's sarcoma-associated herpesvirus (KSHV), also known as human herpesvirus 8 (HHV-8), is a cancer-related human virus, classified as a member of the Gammaherpesvirinae subfamily. We report here the construction of a dual fluorescent-tagged KSHV genome (BAC16-mCherry-ORF45), which constitutively expresses green fluorescent protein (GFP) and contains the tegument multifunctional ORF45 protein as a fusion protein with monomeric Cherry fluorescent protein (mCherry). We confirmed that this virus is properly expressed and correctly replicates and that the mCherry-ORF45 protein is incorporated into the virions. Using this labeled virus, we describe the dynamics of mCherry-ORF45 expression and localization in newly infected cells as well as in latently infected cells undergoing lytic induction and show that mCherry can be used to monitor cells undergoing the lytic viral cycle. This virus is likely to enable future studies monitoring the dynamics of viral trafficking and tegumentation during viral ingress and egress.

Original languageEnglish
Pages (from-to)12839-12852
Number of pages14
JournalJournal of Virology
Volume88
Issue number21
DOIs
StatePublished - Nov 2014

Bibliographical note

Publisher Copyright:
© 2014, American Society for Microbiology.

Funding

FundersFunder number
National Institutes of Health
National Cancer InstituteP01CA180779

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