Jan 29, 2016

Cell re-entry assays do not support models of pathogen- independent translocation of AvrM and AVR3a effectors into plant cells

BioRxiv : the Preprint Server for Biology
Benjamin PetreSophien Kamoun

Abstract

The cell re-entry assay is widely used to evaluate pathogen effector protein uptake into plant cells. The assay is based on the premise that effector proteins secreted out of a leaf cell would translocate back into the cytosol of the same cell via a yet unknown host-derived uptake mechanism. Here, we critically assess this assay by expressing domains of the effector proteins AvrM-A of Melampsora lini and AVR3a of Phytophthora infestans fused to a signal peptide and fluorescent proteins in Nicotiana benthamiana. We found that the secreted fusion proteins do not re-enter plant cells from the apoplast and that the assay is prone to false-positives. We therefore emit a cautionary note on the use of the cell re-entry assay for protein trafficking studies.

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Mentioned in this Paper

Study
Pathogenic Organism
Uptake
Protein Import
Evaluation
Fluorescent stain
Nuclear Translocation
Protein Transport
Enzyme Multiplied Immunoassay Technique
Intracellular Protein Transport

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