RT Journal Article
SR Electronic
T1 Cell re-entry assays do not support models of pathogen-independent translocation of AvrM and AVR3a effectors into plant cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 038232
DO 10.1101/038232
A1 Benjamin Petre
A1 Michaela Kopischke
A1 Alexandre Evrard
A1 Silke Robatzek
A1 Sophien Kamoun
YR 2016
UL http://biorxiv.org/content/early/2016/01/29/038232.abstract
AB 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.