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PMID:21209103
| Citation |
Bamunusinghe, D, Seo, JK and Rao, AL (2011) Subcellular localization and rearrangement of endoplasmic reticulum by Brome mosaic virus capsid protein. J. Virol. 85:2953-63 |
|---|---|
| Abstract |
Genome packaging in the plant-infecting Brome mosaic virus (BMV), a member of the alphavirus-like superfamily, as well as in other positive-strand RNA viruses pathogenic to humans (e.g., poliovirus) and animals (e.g., Flock House virus), is functionally coupled to replication. Although the subcellular localization site of BMV replication has been identified, that of the capsid protein (CP) has remained elusive. In this study, the application of immunofluorescence confocal microscopy to Nicotiana benthamiana leaves expressing replication-derived BMV CP as a green fluorescent protein (GFP) fusion, in conjunction with antibodies to the CP and double-stranded RNA, a presumed marker of RNA replication, revealed that the subcellular localization sites of replication and CP overlap. Our temporal analysis by transmission electron microscopy of ultrastructural modifications induced in BMV-infected N. benthamiana leaves revealed a reticulovesicular network of modified endoplasmic reticulum (ER) incorporating large assemblies of vesicles derived from ER accumulated in the cytoplasm during BMV infection. Additionally, for the first time, we have found by ectopic expression experiments that BMV CP itself has the intrinsic property of modifying ER to induce vesicles similar to those present in BMV infections. The significance of CP-induced vesicles in relation to CP-organized viral functions that are linked to replication-coupled packaging is discussed. |
| Links |
PubMed PMC3067956 Online version:10.1128/JVI.02020-10 |
| Keywords |
Bromovirus/physiology; Capsid Proteins/genetics; Capsid Proteins/metabolism; Endoplasmic Reticulum/metabolism; Endoplasmic Reticulum/virology; Genes, Reporter; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; Microscopy, Confocal; Microscopy, Electron, Transmission; Microscopy, Fluorescence; Plant Leaves/virology; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Tobacco/virology; Virus Assembly |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0044165: host cell endoplasmic reticulum |
ECO:0000314: |
C |
Capsid protein was fused with GFP, and to determine its sub-cellular location, immuno-fluorescence using primary antibodies against two ER markers, namely,KDEL motif or BiP, was used. This permitted dual localization of GFP and each ER marker protein by IFCM. Figure 1C |
complete | ||||
|
part_of |
GO:0044165: host cell endoplasmic reticulum |
ECO:0000314: direct assay evidence used in manual assertion |
C |
Seeded From UniProt |
complete | |||
Notes
See also
References
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