ARA7(Q69L) expression in transgenic Arabidopsis cells induces the formation of enlarged multivesicular bodies
| Autor: | Liwen Jiang, Yi Cai, Takashi Ueda, Tianran Jia, Junqi Wang, Yong Cui, Yu Ding, Akihiko Nakano, Caiji Gao |
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| Rok vydání: | 2013 |
| Předmět: |
Hot Temperature
homotypic fusion Physiology pre-vacuolar compartment Recombinant Fusion Proteins Green Fluorescent Proteins Arabidopsis Mutation Missense Plant Science Biology multivesicular body Green fluorescent protein symbols.namesake Live cell imaging Arabidopsis thaliana ARA7(Q69L) transgenic Arabidopsis cells Arabidopsis Proteins Multivesicular Bodies Immunogold labelling Golgi apparatus Plants Genetically Modified biology.organism_classification Cell biology Transport protein Protein Transport MVB enlargement rab GTP-Binding Proteins Vacuolar transport symbols Research Paper |
| Zdroj: | Journal of Experimental Botany |
| ISSN: | 1460-2431 0022-0957 |
| DOI: | 10.1093/jxb/ert125 |
| Popis: | Arabidopsis thaliana ARA7 (AtRabF2b), a member of the plant Rab5 small GTPases functioning in the vacuolar transport pathway, localizes to pre-vacuolar compartments (PVCs), known as multivesicular bodies (MVBs) in plant cells. Overexpression of the constitutively active GTP-bound mutant of ARA7, ARA7(Q69L), induces the formation of large ring-like structures (1-2 µm in diameter). To better understand the biology of these ARA7(Q69L)-induced ring-like structures, transgenic Arabidopsis cell lines expressing ARA7(Q69L) tagged with green fluorescent protein (GFP) under the control of a heat shock-inducible promoter were generated. In these transgenic cells, robust ring-like structures were formed after 4 h of heat shock induction. Transient co-expression, confocal imaging, and immunogold electron microscopy (immunogold-EM) experiments demonstrated that these GFP-ARA7(Q69L)-labelled ring-like structures were distinct from the Golgi apparatus and trans-Golgi network, but were labelled with an antibody against an MVB marker protein. In addition, live cell imaging and detailed EM analysis showed that the GFP-ARA7(Q69L)-induced spherical structures originated from the homotypic fusion of MVBs. In summary, it was demonstrated that GFP-ARA7(Q69L) expression is an efficient tool for studying PVC/MVB-mediated protein trafficking and vacuolar degradation in plant cells. |
| Databáze: | OpenAIRE |
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