Human Cytomegalovirus Interactions with the Basement Membrane Protein Nidogen 1.

Abstract

In 2000, we reported that human cytomegalovirus (HCMV) induced specific damage on chromosome 1. The capacity of the virus to induce DNA breaks indicated potent interaction between viral proteins and these loci. We have fine mapped the 1q42 breaksite. Transcriptional analysis of genes encoded in close proximity revealed virus-induced downregulation of a single gene, nidogen 1 (NID1). Beginning between 12 and 24 hours postinfection (hpi) and continuing throughout infection, steady-state (ss) NID1 protein levels were decreased in whole-cell lysates and secreted supernatants of human foreskin fibroblasts. Addition of the proteasomal inhibitor MG132 to culture medium stabilized NID1 in virus-infected cells, implicating infection-activated proteasomal degradation of NID1. Targeting of NID1 via two separate pathways highlighted the virus' emphasis on NID1 elimination. NID1 is an important basement membrane protein secreted by many cell types, including the endothelial cells (ECs) lining the vasculature. We found that ss NID1 was also reduced in infected ECs and hypothesized that virus-induced removal of NID1 might offer HCMV a means of increased distribution throughout the host. Supporting this idea, transmigration assays of THP-1 cells seeded onto NID1-knockout (KO) EC monolayers demonstrated increased transmigration. NID1 is expressed widely in the developing fetal central and peripheral nervous systems (CNS and PNS) and is important for neuronal migration and neural network excitability and plasticity and regulates Schwann cell proliferation, migration, and myelin production. We found that NID1 expression was dramatically decreased in clinical samples of infected temporal bones. While potentially beneficial for virus dissemination, HCMV-induced elimination of NID1 may underlie negative ramifications to the infected fetus.IMPORTANCE We have found that HCMV infection promotes the elimination of the developmentally important basement membrane protein nidogen 1 (NID1) from its host. The virus both decreased transcription and induced degradation of expressed protein. Endothelial cell (EC) secretion of basement membrane proteins is critical for vascular wall integrity, and infection equivalently affected NID1 protein levels in these cells. We found that the absence of NID1 in an EC monolayer allowed increased transmigration of monocytes equivalent to that observed after infection of ECs. The importance of NID1 in development has been well documented. We found that NID1 protein was dramatically reduced in infected inner ear clinical samples. We believe that HCMV's attack on host NID1 favors viral dissemination at the cost of negative developmental ramifications in the infected fetus.

• Authors: Man I Kuan, Hannah K Jaeger, Onesmo B Balemba, John M O'Dowd, Deborah Duricka, Holger Hannemann, Emmerentia Marx, Natacha Teissier, Liliana Gabrielli, Maria Paola Bonasoni, Elizabeth M Keithley, Elizabeth A Fortunato
• Journal: Journal of virology (J Virol) Vol. 95 Issue 3 (01 13 2021) ISSN: 1098-5514 [Electronic] United States
• PMID: 33177203 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2021 American Society for Microbiology.

Chemical References

• Membrane Glycoproteins
• nidogen

Topics

• Basement Membrane (metabolism)
• Cell Movement
• Cytomegalovirus (physiology)
• Cytomegalovirus Infections (metabolism, pathology, virology)
• Endothelium, Vascular (metabolism, virology)
• Fibroblasts (metabolism, virology)
• Humans
• Membrane Glycoproteins (antagonists & inhibitors, genetics, metabolism)
• Signal Transduction
• Virus Internalization