Characterization of fusion from without induced by herpes simplex virus.
The process of fusion from without (FFWO) induced by herpes simplex virus (HSV) was analyzed by using various inhibitors and compared to fusion from within (FFWI). The fate of certain elements of the cytoskeleton after FFWO was also investigated. Our experiments demonstrate FFWO as a very suitable system for study of early virus-cell interactions. Zn++ ions proved inhibitory for penetration whilst pretreatment of cells with Ca++ ions before infection enhanced FFWO activity. Dissociation of penetration from the fusion process itself was possible by use of Zn++ ions, low pH-treatment and antiserum on the one hand and N-ethylmaleimide and cytochalasin D on the other. Penetration itself needs only 6 min or less to proceed. FFWO is independent of inhibitors of glycosylation (tunicamycin) and intracellular vesicular traffic (monensin), protein-synthesis (cycloheximide) and energy-delivery (2.4 dinitrophenol and Na-azide). Analyzed strains of HSV-1 and -2 producing FFWI could be subgrouped into three categories: Strain ANG with high, strain HFEM and Lux with low and strains IES, Len, MP, US with no FFWO activity. The results of these experiments indicate that the property of FFWO is not purely a consequence of the number of PFU but depends on certain inherent properties of the virus particles. Addition of heparin as well as treatment of cells with heparitinase effectively prevented FFWO, indicating identical virus receptors for entrance of virus into cells and FFWO. During our studies several calf sera were found to inhibit FFWO-activity. Inhibition of FFWO by a glycoconjugate (ferritin coupled with oleic acid) indicates specific stereochemical hindrance of FFWO by this compound. Shortly after FFWO the actin filaments rearrange to form long fibres and surface fibronectin is being lost from the cell membrane.
Walev
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I
Division of Experimental Virology, University of Mainz, Federal Republic of Germany.
Wollert
K C
KC
Weise
K
K
Falke
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D
D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D002459:Cell Fusion; D002462:Cell Membrane / Q000382:microbiology*; D003599:Cytoskeleton / Q000382:microbiology; D005033:Ethylmaleimide / Q000494:pharmacology; D006001:Glycoconjugates / Q000494:pharmacology; D037102:Lectins / Q000494:pharmacology; D008055:Lipids / Q000494:pharmacology; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011991:Receptors, Virus / Q000187:drug effects; D018139:Simplexvirus / Q000502:physiology*; D013045:Species Specificity; D014709:Vero Cells
D002413:Cations, Divalent; D006001:Glycoconjugates; D037102:Lectins; D008055:Lipids; D011480:Protease Inhibitors; D011991:Receptors, Virus; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase; D005033:Ethylmaleimide