Deletions into an NH2-terminal hydrophobic domain result in secretion of rotavirus VP7, a resident endoplasmic reticulum membrane glycoprotein
| Autor: | Paul H. Atkinson, G W Both, F Sato, M. S. Poruchynsky, C Tyndall, A R Bellamy |
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| Rok vydání: | 1985 |
| Předmět: |
Rotavirus
viruses Fluorescent Antibody Technique Biology Protein Sorting Signals Endoplasmic Reticulum symbols.namesake chemistry.chemical_compound Structure-Activity Relationship Viral Proteins Chlorocebus aethiops Morphogenesis Animals Secretion Amino Acid Sequence Integral membrane protein Glycoproteins chemistry.chemical_classification Endoplasmic reticulum membrane Endoplasmic reticulum Membrane Proteins STIM1 Cell Biology Tunicamycin Articles Golgi apparatus Molecular biology Cell Compartmentation Molecular Weight Microscopy Electron Hexosaminidases chemistry Solubility symbols Glycoprotein Protein Processing Post-Translational |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 0021-9525 |
| Popis: | Rotavirus, a non-enveloped reovirus, buds into the rough endoplasmic reticulum and transiently acquires a membrane. The structural glycoprotein, VP7, a 38-kD integral membrane protein of the endoplasmic reticulum (ER), presumably transfers to virus in this process. The gene for VP7 potentially encodes a protein of 326 amino acids which has two tandem hydrophobic domains at the NH2-terminal, each preceded by an in-frame ATG codon. A series of deletion mutants constructed from a full-length cDNA clone of the Simian 11 rotavirus VP7 gene were expressed in COS 7 cells. Products from wild-type, and mutants which did not affect the second hydrophobic domain of VP7, were localized by immunofluorescence to elements of the ER only. However, deletions affecting the second hydrophobic domain (mutants 42-61, 43-61, 47-61) showed immunofluorescent localization of VP7 which coincided with that of wheat germ agglutinin, indicating transport to the Golgi apparatus. Immunoprecipitable wild-type protein, or an altered protein lacking the first hydrophobic sequence, remained intracellular and endo-beta-N-acetylglucosaminidase H sensitive. In contrast, products of mutants 42-61, 43-61, and 47-61 were transported from the ER, and secreted. Glycosylation of the secreted molecules was inhibited by tunicamycin, resistant to endo-beta-N-acetylglucosaminidase H digestion and therefore of the N-linked complex type. An unglycosylated version of VP7 was also secreted. We suggest that the second hydrophobic domain contributes to a positive signal for ER location and a membrane anchor function. Secretion of the mutant glycoprotein implies that transport can be constitutive with the destination being dictated by an overriding compartmentalization signal. |
| Databáze: | OpenAIRE |
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