Characterization of a highly glycosylated form of the human cytomegalovirus HLA class I homologue gpUL18.
Human cytomegalovirus (HCMV) gpUL18 is a HLA class I (HLA-I) homologue with high affinity for the inhibitory receptor LIR-1/ILT2. The previously described 67 kDa form of gpUL18 is shown here to be sensitive to endoglycosidase-H (EndoH). A novel form of gpUL18 with a molecular mass of approximately 160 kDa and resistance to EndoH was identified in cells infected with HCMV strain AD169 or the low passage HCMV isolates Merlin and Toledo. The 67 kDa EndoH-sensitive gpUL18 glycoform was detected earlier in a productive infection (from 24 h post-infection) than the slower-migrating EndoH-resistant glycoform (from 72 h post-infection). Deletion of the US2-US11 region from the HCMV genome was associated with a substantial up-regulation of endogenous HLA-I in infected cells, but had no obvious effect on the gpUL18 expression pattern. Vaccinia virus and adenovirus vectors were used to further analyse gpUL18 expression. Depending on the delivery vector system, differences in the electrophoretic motility of the EndoH-resistant >105 kDa form of gpUL18, but not the EndoH-sensitive 67 kDa form, were observed; post-translational modification of the higher molecular mass glycoform appears to be influenced by active virus infection and vector delivery. The EndoH-sensitive 67 kDa gpUL18 had a rapid turnover, while the maturation to the EndoH-resistant >105 kDa form was relatively slow and inefficient. However, synthesis of the EndoH-resistant >105 kDa form was enhanced with elevated levels of beta2-microglobulin. When expressed by using an adenovirus vector, both the EndoH-sensitive 67 kDa and the EndoH-resistant >105 kDa gpUL18 forms could be detected on the cell surface.
Griffin
Cora
C
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
Wang
Eddie C Y
ECY
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
McSharry
Brian P
BP
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
Rickards
Carole
C
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
Browne
Helena
H
Division of Virology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
Wilkinson
Gavin W G
GWG
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
Tomasec
Peter
P
Section of Infection and Immunity, Cardiff University, Tenovus Building, Heath Park, Cardiff CF14 4XX, UK.
D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003587:Cytomegalovirus / Q000737:chemistry* / Q000235:genetics; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000737:chemistry; D006801:Humans
D036022:Capsid Proteins; D006023:Glycoproteins; D015395:Histocompatibility Antigens Class I; C079750:VP23 protein, Human herpesvirus 1