Membrane topology of the vaccinia virus A17L envelope protein.
The formation of a lipoprotein membrane within specialized areas of the cytoplasm is the first visible step in poxvirus morphogenesis. The A17L viral protein, an essential nonglycosylated membrane component, was predicted to have four centrally located alpha-helical membrane-spanning domains. The gene was expressed as a 23-kDa protein in a cell-free transcription/translation system containing canine pancreatic microsomes. The N- and C-terminal ends of the membrane-associated protein were susceptible to proteinase digestion, whereas the central region was resistant, consistent with a model in which the first and fourth hydrophobic domains are membrane spanning. This topology was supported by the sizes of the major proteinase-resistant membrane-associated products of genes containing one or more deleted hydrophobic domains and by evidence that the C-terminus was intraluminal and glycosylated on deletion of the second, third, and fourth domains, the third and fourth domains, or just the fourth domain. Moreover, glycosylation also occurred when an N-glycosylation site was introduced into the second hydrophobic domain of the full-length A17L protein. The data indicated a predominant topology in which the N- and C-termini are cytoplasmic, the first and fourth hydrophobic domains span the microsomal membrane, and the second and third hydrophobic domains are intraluminal. This arrangement has important implications for interactions of the A17L protein with other membrane components.
Betakova
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T
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892-0445, USA.
Wolffe
E J
EJ
Moss
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B
D000595:Amino Acid Sequence; D000818:Animals; D004285:Dogs; D015967:Gene Expression Regulation, Viral; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D017384:Sequence Deletion; D014616:Vaccinia virus / Q000737:chemistry / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
C081257:A17L protein, Vaccinia virus; D008565:Membrane Proteins; D014759:Viral Envelope Proteins