title: Insertional mutations in herpes simplex virus type 1 gL identify functional domains for association with gH and for membrane fusion.
abstract: Glycoprotein L (gL) is one of four glycoproteins required for the entry of herpes simplex virus (HSV) into cells and for virus-induced cell fusion. This glycoprotein oligomerizes with gH to form a membrane-bound heterodimer but can be secreted when expressed without gH. Twelve unique gL linker-insertion mutants were generated to identify regions critical for gH binding and gH/gL processing and regions essential for cell fusion and viral entry. All gL mutants were detected on the cell surface in the absence of gH, suggesting incomplete cleavage of the signal peptide or the presence of a cell surface receptor for secreted gL. Coexpression with gH enhanced the levels of cell surface gL detected by antibodies for all gL mutants except those that were defective in their interactions with gH. Two insertions into a conserved region of gL abrogated the binding of gL to gH and prevented gH expression on the cell surface. Three other insertions reduced the cell surface expression of gH and/or altered the properties of gH/gL heterodimers. Altered or absent interaction of gL with gH was correlated with reduced or absent cell fusion activity and impaired complementation of virion infectivity. These results identify a conserved domain of gL that is critical for its binding to gH and two noncontiguous regions of gL, one of which contains the conserved domain, that are critical for the gH/gL complex to perform its role in membrane fusion.
authors.affiliation: Department of Microbiology-Immunology, The Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Fan
authors.forename: Erick
authors.initials: E
authors.lastname: Lin
authors.forename: Patricia G
authors.initials: PG
authors.lastname: Spear
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D019281:Dimerization; D018259:Herpesvirus 1, Human / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000235:genetics*; D017434:Protein Structure, Tertiary / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000502:physiology; D016662:Virus Integration / Q000235:genetics; D053586:Virus Internalization
chemical_list: D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1; C077407:glycoprotein L, Human herpesvirus 1