title: Gammaherpesvirus 68 infection of endothelial cells requires both host autophagy genes and viral oncogenes for optimal survival and persistence.
abstract: Gammaherpesvirus-associated neoplasms include tumors of lymphocytes, epithelial cells, and endothelial cells (ECs). We previously showed that, unlike most cell types, ECs survive productive gammaherpesvirus 68 (γHV68) infection and achieve anchorage-independent growth, providing a cellular reservoir for viral persistence. Here, we demonstrated autophagy in infected ECs by analysis of LC3 localization and protein modification and that infected ECs progress through the autophagosome pathway by LC3 dual fluorescence and p62 analysis. We demonstrate that pharmacologic autophagy induction results in increased survival of infected ECs and, conversely, that autophagy inhibition results in death of infected EC survivors. Furthermore, we identified two viral oncogenes, v-cyclin and v-Bcl2, that are critical to EC survival and that modify EC proliferation and survival during infection-induced autophagy. We found that these viral oncogenes can also facilitate survival of substrate detachment in the absence of viral infection. Autophagy affords cells the opportunity to recover from stressful conditions, and consistent with this, the altered phenotype of surviving infected ECs was reversible. Finally, we demonstrated that knockdown of critical autophagy genes completely abrogated EC survival. This study reveals a viral mechanism which usurps the autophagic machinery to promote viral persistence within nonadherent ECs, with the potential for recovery of infected ECs at a distant site upon disruption of virus replication.
authors.affiliation: Department of Microbiology, University of Colorado Denver School of Medicine, 12800 E. 19th Avenue, Aurora, CO 80045, USA.
authors.forename: Andrea Luísa
authors.initials: AL
authors.lastname: Suárez
authors.forename: Raymond
authors.initials: R
authors.lastname: Kong
authors.forename: Tad
authors.initials: T
authors.lastname: George
authors.forename: Liqiang
authors.initials: L
authors.lastname: He
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Yue
authors.forename: Linda Faye
authors.initials: LF
authors.lastname: van Dyk
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000473:pathology / Q000502:physiology / Q000821:virology*; D018142:Gammaherpesvirinae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology*; D006566:Herpesviridae Infections / Q000473:pathology / Q000821:virology*; D008168:Lung / Q000166:cytology; D051379:Mice; D008822:Mice, Transgenic; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D009857:Oncogenes / Q000235:genetics* / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D015513:Oncogene Proteins; D014764:Viral Proteins