Reconstitution of the Kaposi's sarcoma-associated herpesvirus nuclear egress complex and formation of nuclear membrane vesicles by coexpression of ORF67 and ORF69 gene products.
The Kaposi's sarcoma-associated herpesvirus nuclear egress complex is composed of two proteins, ORF67 and ORF69. In this study, we have recapitulated the KSHV complex by coexpression of these two proteins in insect cells using expression from recombinant baculoviruses. The proteins form a complex at the nuclear membrane as judged by live-cell analysis of protein fusions tagged with green fluorescent protein (GFP) and mCherry. Ultrastructural analysis of infected cells showed that ORF67 expression results in reduplication of the nuclear membrane. When the two proteins are expressed together, numerous virion-size nuclear membrane-derived vesicles were evident at the nuclear margins.
Viral Oncology Program, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Johns Hopkins University, School of Medicine, Baltimore, Maryland, USA. pdesai@jhmi.edu
Prashant J
PJ
Desai
Erin N
EN
Pryce
Brandon W
BW
Henson
Eric M
EM
Luitweiler
Jonathan
J
Cothran
D000818:Animals; D002460:Cell Line; D022162:Cytoplasmic Vesicles / Q000821:virology*; D006566:Herpesviridae Infections / Q000821:virology*; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D009685:Nuclear Envelope / Q000821:virology*; D011485:Protein Binding; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D057074:Virus Release
D014764:Viral Proteins; C529434:p33 protein, Kaposi's sarcoma-associated herpesvirus