Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.795839
Title: Herpes simplex virus-induced glycoproteins : mapping and characterisation of g92K and gE
Author: Hope, Ralph Graham
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1986
Availability of Full Text:
Access from EThOS:
Access from Institution:
Abstract:
Cells infected with HSV-1 (strain 17 syn+) or HSV-2 (strain HG52) incorporated inorganic sulphate into polypeptides which comigrated on SDS polyacrylamide gels with virus-induced glycoproteins. The major sulphated glycoprotein was glycoprotein E. In addition, inorganic sulphate was incorporated into glycoprotein D and HSV-1 glycoproteins B, C and Y. Incorporation of sulphate label into HSV-2 glycoproteins B/C was occasionally observed. The addition of inorganic sulphate occurs late during glycoprotein maturation. Analysis of intracellular sulphated polypeptides using intertypic recombinants mapped glycoprotein E to between 0.886 and 0.935 map units (mu) of the HSV genome. Studies from other laboratories had established the types of oligosaccharide linkages on glycoproteins B, C and D. Experiments are reported in this thesis in which the drug tunicamycin was used to investigate the nature of the linkage of oligosaccharides to HSV-1 induced glycoproteins E and Y and the nature of the sulphate linkage to all the recognised HSV-1 glycoproteins. Synthesis of both gE-1 and gY-1 was inhibited by the drug, suggesting they contain N-linked oligosaccharides. Tunicamycin also inhibited the incorporation of inorganic sulphate into all HSV-1 glycoproteins although reduced amounts of sulphate could be detected in an abnormal form of gE-1. These results suggest that most inorganic sulphate appears to be attached to N-linked oligosaccharides but for gE-1 some may be attached to the polypeptide backbone or to O-linked oligosaccharides. Major sulphated species of apparent MW 32000, 34000 and 35000 were secreted from cells infected with 17 syn+. In addition, sulphated polypeptides which migrated in the vicinity of glycoprotein D were secreted from cells infected with 17 syn+. These species were subsequently shown by tryptic peptide fingerprinting to be encoded by the gene encoding glycoprotein E. Furthermore, over 95% of the total amount of 32000, 34000 and 35000 polypeptides synthesised was secreted. The secreted proteins are produced when infected cells are incubated in the presence of serum but are not produced in its absence demonstrating that a serum component is responsible for their generation. Unlike gE, the secreted proteins do not possess affinity for the Fc end of IgG. Evidence is presented showing that the 92000-dalton glycoprotein (g92K) induced by HSV-2 has properties distinct from those assigned to any other HSV-2 glycoproteins. First, the carbohydrate composition and extent of sulphation differ from those of glycoproteins D and E. Second, two clonally unrelated monoclonal antibodies, AP1 and LP5, shown to specifically immunoprecipitate g92K, do not react with any of the known processed forms of glycoproteins B, C, D and E. Third, by using HSV-1 x HSV-2 intertypic recombinants g92K was shown to map in the short region of the HSV genome (0.846 - 0.924). Fourth, the intertypic recombinant R12-1 which did not induce g92K, induced HSV-2 gE and an altered gD, providing genetic evidence that g92K is encoded, at least in part, by a different region of the genome from that encoding gE.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.795839  DOI: Not available
Share: