ABSTRACTEndothelial cells (ECs) are a critical target of viruses, and infection of the endothelium represents a defining point in viral pathogenesis. Human cytomegalovirus (HCMV), the prototypical betaherpesvirus, encodes proteins specialized for entry into ECs and delivery of the genome to the nuclei of ECs. Virus strains competent to enter ECs replicate with differing efficiencies, suggesting that the virus encodes genes for postentry tropism in ECs. We previously reported a specific requirement for theUL133/8locus of HCMV for replication in ECs. TheUL133/8locus harbors four genes:UL133,UL135,UL136, andUL138. In this study, we find that whileUL133andUL138are dispensable for replication in ECs, bothUL135andUL136are important. These genes are not required for virus entry or the expression of viral genes. The phenotypes associated with disruption of either gene reflect phenotypes observed for theUL133/8NULLvirus, which lacks the entireUL133/8locus, but are largely distinct from one another. Viruses lackingUL135fail to properly envelop capsids in the cytoplasm, produce fewer dense bodies (DB) than the wild-type (WT) virus, and are unable to incorporate viral products into multivesicular bodies (MVB). Viruses lackingUL136also fail to properly envelop virions and produce larger dense bodies than the WT virus. Our results indicate roles for theUL135andUL136proteins in commandeering host membrane-trafficking pathways for virus maturation.UL135andUL136represent the first HCMV genes crucial for early- to late-stage tropism in ECs.IMPORTANCEHuman cytomegalovirus (HCMV) persists in the majority of the world's population. While typically asymptomatic in healthy hosts, HCMV can cause significant morbidity and mortality in immunocompromised or naïve individuals, particularly transplant patients and patients with congenital infections, respectively. Lifelong persistence of the virus may also contribute to age-related pathologies, such as vascular disease. One aspect of HCMV infection contributing to complex and varied pathogenesis is the diverse array of cell types that this virus infects in the host. The vascular endothelium is a particularly important target of infection, contributing to viral dissemination and likely leading to CMV complications following transplantation. In this work, we identify two viral gene products required for postentry tropism in endothelial cells. Identifying tropism factors required for replication in critical cell targets of infection is important for the development of strategies to restrict virus replication.
Legionella pneumophila LegC7 effector protein drives aberrant ER:endosome fusion in yeast