Insertion mutations in herpes simplex virus 1 glycoprotein H reduce cell surface expression, slow the rate of cell fusion, or abrogate functions in cell fusion and viral entry.
Of the four required herpes simplex virus (HSV) entry glycoproteins, the precise role of gH-gL in fusion remains the most elusive. The heterodimer gH-gL has been proposed to mediate hemifusion after the interaction of another required glycoprotein, gD, with a receptor. To identify functional domains of HSV-1 gH, we generated 22 randomized linker-insertion mutants. Analyses of 22 gH mutants revealed that gH is relatively tolerant of insertion mutations, as 15 of 22 mutants permitted normal processing and transport of gH-gL to the cell surface. gH mutants that were not expressed well at the cell surface did not function in fusion or viral entry. The screening of gH mutants for function revealed the following: (i) for wild-type gH and some gH mutants, fusion with nectin-1-expressing target cells occurred more rapidly than with herpesvirus entry mediator (HVEM)-expressing target cells; (ii) some gH mutants reduced the rate of cell fusion without abrogating fusion completely, indicating that gH may play a role in governing the kinetics of fusion and may be responsible for a rate-limiting first stage in HSV-1 fusion; and (iii) only one gH mutant, located within the short cytoplasmic tail, completely abrogated function, indicating that the gH cytoplasmic tail is crucial for cell fusion and viral infectivity.
Jackson
Julia O
JO
Department of Microbiology and Immunology, the Feinberg School of Medicine, Northwestern University, 303 East Chicago Avenue, Ward 6-231, Chicago, IL 60611, USA.
Lin
Erick
E
Spear
Patricia G
PG
Longnecker
Richard
R
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005816:Genetic Complementation Test; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D016254:Mutagenesis, Insertional; D017434:Protein Structure, Tertiary; D053240:Receptors, Tumor Necrosis Factor, Member 14 / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D053586:Virus Internalization
D053240:Receptors, Tumor Necrosis Factor, Member 14; D011994:Recombinant Proteins; C506028:TNFRSF14 protein, human; D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1