Necroptosis-based CRISPR knockout screen reveals Neuropilin-1 as a critical host factor for early stages of murine cytomegalovirus infection

• Type of Material: Article
• Authors: Rebecca K. Lane, University of Texas Health Science Center at San Antonio; Hongyuan Guo, Georgia State University; Amanda D. Fisher, University of Texas Health Science Center at San Antonio; Jonathan Diep, Stanford University; Zhao Lai, University of Texas Health Science Center at San Antonio; Yidong Chen, University of Texas Health Science Center at San Antonio; Jason W. Upton, Auburn University; Jan Carette, Stanford University; Edward Mocarski, Emory University; William Kaiser, University of Texas Health Science Center at San Antonio
• Language: English
• Date: 2020-08-18
• Publisher: National Academy of Sciences
• Publication Version: Final Published Version
• Copyright Statement: © 2020 National Academy of Science. All rights reserved.
• Final Published Version (URL): http://dx.doi.org/10.1073/pnas.1921315117
• Title of Journal or Parent Work: Proceedings of the National Academy of Sciences of the United States
• Volume: 117
• Issue: 33
• Start Page: 20109
• End Page: 20116
• Grant/Funding Information: Data were generated in the Flow Cytometry Shared Resource Facility, which is supported by University of Texas Health San Antonio, Grant NIH-National Cancer Institute (NCI) P30 CA054174-20 (Cancer Therapy & Research Center at UTHSCSA), UL1 TR001120 (Clinical and Translational Science Award), and the Genome Sequencing Facility, which is supported by UTHSCSA, NIH-NCI P30 CA054174 (Cancer Center Support Grant to UTHSCSA), and Cancer Prevention and Research Institute of Texas Core Facility Award (RP160732).; This work was also supported by NIH grants R01 AI141970 to J.C., DP5 OD012198 to W.J.K., and the Craniofacial Oral-biology Student Training in Academic Research fellowship award (T32DE014318-16) and National Institute of Dental and Craniofacial Research F31 predoctoral fellowship F31DE029395 to R.K.L.
• Keywords: necroptosis; neuropilin; Heparan sulfate; Entry; Cells; Growth; Virus; Science & Technology; Science & Technology - Other Topics; cytomegolovirus; Structural basis; CRISPR; Necrosis; Multidisciplinary Sciences; Binding; Activation
• Research Categories: Biology, Microbiology; Health Sciences, Immunology; Biology, Molecular; Biology, Virology

Abstract

Herpesviruses are ubiquitous human pathogens that cause a wide range of health complications. Currently, there is an incomplete understanding of cellular factors that contribute to herpesvirus infection. Here, we report an antiviral necroptosis-based genetic screen to identify novel host cell factors required for infection with the β-herpesvirus murine cytomegalovirus (MCMV). Our genomewide CRISPR-based screen harnessed the capacity of herpesvirus mutants that trigger antiviral necroptotic cell death upon early viral gene expression. Vascular endothelial growth factor (VEGF) and semaphorin-binding receptor Neuropilin-1 (Nrp-1) emerge as crucial determinants of MCMV infection. We find that elimination of Nrp-1 impairs early viral gene expression and reduces infection rates in endothelial cells, fibroblasts, and macrophages. Furthermore, preincubation of virus with soluble Nrp-1 dramatically inhibits infection by reducing virus attachment. Thus, Nrp-1 is a key determinant of the initial phase of MCMV infection.