Glycosylation of human papillomavirus type 16 L1 protein.
We examined glycosylation of the L1 capsid protein of human papillomavirus type 16, using HPV16 L1 protein expressed from various recombinant vaccinia viruses in CV-1 and HaCaT cells. A minority of L1 protein was N-glycosylated, and all four potential N-glycosylation sites appeared to be used. Glycosylation was of the high-mannose type, as shown by reactivity with biotin-labeled Concanavalin A and by exoglycosidase digestions. A series of mutant L1 proteins were used to establish that an N-terminal hydrophobic sequence, common to all sequenced papillomavirus L1 capsid proteins, was a major determinant of the proportion of L1 protein glycosylated, whereas C-terminus nuclear localization signal sequences were unimportant. Subcellular localization studies showed that whereas the majority of L1 protein was found in the cell nucleus, glycosylated L1 was retained in the endoplasmic reticulum and was neither exported from the cell nor translocated to the cell membrane or the cell nucleus. We conclude that glycosylated L1 is unlikely to be an important component of the papillomavirus virion, a finding of importance for the design of papillomavirus-specific vaccines.
Zhou
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Papillomavirus Research Unit, Princess Alexandra Hospital, Brisbane Qld., Australia.
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D000118:Acetylglucosaminidase / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D036022:Capsid Proteins; D002451:Cell Compartmentation; D004252:DNA Mutational Analysis; D005455:Fluorescent Antibody Technique; D006031:Glycosylation / Q000187:drug effects; D037102:Lectins; D008358:Mannose; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D027383:Papillomaviridae / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology; D014616:Vaccinia virus / Q000235:genetics
D036022:Capsid Proteins; D037102:Lectins; D009844:Oligosaccharides; D009856:Oncogene Proteins, Viral; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D014415:Tunicamycin; C063317:L1 protein, Human papillomavirus type 16; D000118:Acetylglucosaminidase; D008358:Mannose