Application of phiLOV2.1 as a fluorescent marker for visualization of Agrobacterium effector protein translocation
| Autor: | Mohammad Reza Roushan, Gerard Paul H. van Heusden, Milou A. M. de Zeeuw, Paul J. J. Hooykaas |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Agrobacterium Recombinant Fusion Proteins Nicotiana tabacum Green Fluorescent Proteins 030106 microbiology Saccharomyces cerevisiae Arabidopsis Gene Expression Chromosomal translocation Plant Science Ion Channels Green fluorescent protein 03 medical and health sciences Bacterial Proteins Tobacco Genetics Secretion biology Effector fungi food and beverages Cell Biology Agrobacterium tumefaciens biology.organism_classification Cell biology DNA-Binding Proteins Protein Transport |
| Zdroj: | Plant Journal, 96(3), 685–699 Plant Journal |
| Popis: | Agrobacterium tumefaciens can genetically transform plants by translocating a piece of oncogenic DNA, called T‐DNA, into host cells. Transfer is mediated by a type IV secretion system (T4SS). Besides the T‐DNA, which is transferred in a single‐stranded form and at its 5′ end covalently bound to VirD2, several other effector proteins (VirE2, VirE3, VirD5, and VirF) are translocated into the host cells. The fate and function of the translocated proteins inside the host cell are only partly known. Therefore, several studies were conducted to visualize the translocation of the VirE2 protein. As GFP‐tagged effector proteins are unable to pass the T4SS, other approaches like the split GFP system were used, but these require specific transgenic recipient cells expressing the complementary part of GFP. Here, we investigated whether use can be made of the photostable variant of LOV, phiLOV2.1, to visualize effector protein translocation from Agrobacterium to non‐transgenic yeast and plant cells. We were able to visualize the translocation of all five effector proteins, both to yeast cells, and to cells in Nicotiana tabacum leaves and Arabidopsis thaliana roots. Clear signals were obtained that are easily distinguishable from the background, even in cases in which by comparison the split GFP system did not generate a signal. |
| Databáze: | OpenAIRE |
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