title: Rho GTPases modulate entry of Ebola virus and vesicular stomatitis virus pseudotyped vectors.
abstract: To explore mechanisms of entry for Ebola virus (EBOV) glycoprotein (GP) pseudotyped virions, we used comparative gene analysis to identify genes whose expression correlated with viral transduction. Candidate genes were identified by using EBOV GP pseudotyped virions to transduce human tumor cell lines that had previously been characterized by cDNA microarray. Transduction profiles for each of these cell lines were generated, and a significant positive correlation was observed between RhoC expression and permissivity for EBOV vector transduction. This correlation was not specific for EBOV vector alone as RhoC also correlated highly with transduction of vesicular stomatitis virus GP (VSVG) pseudotyped vector. Levels of RhoC protein in EBOV and VSV permissive and nonpermissive cells were consistent with the cDNA gene array findings. Additionally, vector transduction was elevated in cells that expressed high levels of endogenous RhoC but not RhoA. RhoB and RhoC overexpression significantly increased EBOV GP and VSVG pseudotyped vector transduction but had minimal effect on human immunodeficiency virus (HIV) GP pseudotyped HIV or adeno-associated virus 2 vector entry, indicating that not all virus uptake was enhanced by expression of these molecules. RhoB and RhoC overexpression also significantly enhanced VSV infection. Similarly, overexpression of RhoC led to a significant increase in fusion of EBOV virus-like particles. Finally, ectopic expression of RhoC resulted in increased nonspecific endocytosis of fluorescent dextran and in formation of increased actin stress fibers compared to RhoA-transfected cells, suggesting that RhoC is enhancing macropinocytosis. In total, our studies implicate RhoB and RhoC in enhanced productive entry of some pseudovirions and suggest the involvement of actin-mediated macropinocytosis as a mechanism of uptake of EBOV GP and VSVG pseudotyped viral particles.
authors.affiliation: Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Kathrina
authors.initials: K
authors.lastname: Quinn
authors.forename: Melinda A
authors.initials: MA
authors.lastname: Brindley
authors.forename: Melodie L
authors.initials: ML
authors.lastname: Weller
authors.forename: Nikola
authors.initials: N
authors.lastname: Kaludov
authors.forename: Andrew
authors.initials: A
authors.lastname: Kondratowicz
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Hunt
authors.forename: Patrick L
authors.initials: PL
authors.lastname: Sinn
authors.forename: Paul B
authors.initials: PB
authors.lastname: McCray
authors.forename: Colleen S
authors.initials: CS
authors.lastname: Stein
authors.forename: Beverly L
authors.initials: BL
authors.lastname: Davidson
authors.forename: Ramon
authors.initials: R
authors.lastname: Flick
authors.forename: Robert
authors.initials: R
authors.lastname: Mandell
authors.forename: William
authors.initials: W
authors.lastname: Staplin
authors.forename: Wendy
authors.initials: W
authors.lastname: Maury
authors.forename: John A
authors.initials: JA
authors.lastname: Chiorini
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D016360:Clostridioides difficile; D029043:Ebolavirus / Q000201:enzymology*; D005822:Genetic Vectors; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D008954:Models, Biological; D010957:Plasmids / Q000378:metabolism; D014709:Vero Cells; D018116:Vesiculovirus / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
chemical_list: D020741:rho GTP-Binding Proteins