Trafficking of varicella-zoster virus glycoprotein gI: T(338)-dependent retention in the trans-Golgi network, secretion, and mannose 6-phosphate-inhibitable uptake of the ectodomain.
The trans-Golgi network (TGN) is putatively the site where varicella-zoster virus is enveloped. gE is targeted to the TGN by selective retrieval from the plasmalemma in response to signaling sequences in its endodomain. gI lacks these sequences but forms a complex with gE. We now find that gI is targeted to the TGN and plasma membrane when expressed in Cos-7 cells; nevertheless, surface labeling revealed that gI is not retrieved from the plasma membrane. TGN targeting of gI depended on the T(338) of its endodomain and was lost when T(338) was deleted or mutated to A, S, or D. The endodomain of gI was sufficient, if it contained T(338), to target a fusion protein containing the ectodomain of the human interleukin-2 receptor to the TGN. A truncated protein consisting only of the gI ectodomain was secreted and taken up by nontransfected cells. This uptake of the secreted gI ectodomain was blocked by mannose 6-phosphate. Following cotransfection, both gI and gE were retrieved to the TGN from the plasma membrane in 26.7% of cells, neither gI nor gE was internalized in 18.3%, and gE was retrieved to the TGN while gI remained at the plasma membrane in 55%. We suggest that the T(338) of its endodomain is necessary to retain gI in the TGN; moreover, because gI and gE interact, the signaling sequences of each glycoprotein reinforce one another in ensuring that both glycoproteins are concentrated in the TGN yet remain on the cell surface.
Wang
Z H
ZH
Institute of Human Nutrition, Columbia University College of Physicians and Surgeons, New York, New York 10032, USA.
Gershon
M D
MD
Lungu
O
O
Zhu
Z
Z
Gershon
A A
AA
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001692:Biological Transport; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism*; D014645:Herpesvirus 3, Human / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D008360:Mannosephosphates / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D015375:Receptors, Interleukin-2 / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020539:Sequence Analysis, Protein; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
D008360:Mannosephosphates; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C070916:glycoprotein IV, varicella-zoster virus; C027693:mannose-6-phosphate