title: Agp2p, the plasma membrane transregulator of polyamine uptake, regulates the antifungal activities of the plant defensin NaD1 and other cationic peptides.
abstract: Cationic antifungal peptides (AFPs) act through a variety of mechanisms but share the common feature of interacting with the fungal cell surface. NaD1, a defensin from Nicotiana alata, has potent antifungal activity against a variety of fungi of both hyphal and yeast morphologies. The mechanism of action of NaD1 occurs via three steps: binding to the fungal cell surface, permeabilization of the plasma membrane, and internalization and interaction with intracellular targets to induce fungal cell death. The targets at each of these three stages have yet to be defined. In this study, the screening of a Saccharomyces cerevisiae deletion collection led to the identification of Agp2p as a regulator of the potency of NaD1. Agp2p is a plasma membrane protein that regulates the transport of polyamines and other molecules, many of which carry a positive charge. Cells lacking the agp2 gene were more resistant to NaD1, and this resistance was accompanied by a decreased uptake of defensin. Agp2p senses and regulates the uptake of the polyamine spermidine, and competitive inhibition of the antifungal activity of NaD1 by spermidine was observed in both S. cerevisiae and the plant pathogen Fusarium oxysporum. The resistance of agp2Δ cells to other cationic antifungal peptides and decreased binding of the cationic protein cytochrome c to agp2Δ cells compared to that of wild-type cells have led to a proposed mechanism of resistance whereby the deletion of agp2 leads to an increase in positively charged molecules at the cell surface that repels cationic antifungal peptides.
authors.affiliation: La Trobe Institute for Molecular Science, Melbourne, Victoria, Australia.
authors.forename: Mark R
authors.initials: MR
authors.lastname: Bleackley
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Wiltshire
authors.forename: Francine
authors.initials: F
authors.lastname: Perrine-Walker
authors.forename: Shaily
authors.initials: S
authors.lastname: Vasa
authors.forename: Rhiannon L
authors.initials: RL
authors.lastname: Burns
authors.forename: Nicole L
authors.initials: NL
authors.lastname: van der Weerden
authors.forename: Marilyn A
authors.initials: MA
authors.lastname: Anderson
mesh_terms: D000935:Antifungal Agents / Q000378:metabolism* / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D005434:Flow Cytometry; D005670:Fusarium / Q000187:drug effects / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism* / Q000494:pharmacology; D010940:Plant Proteins / Q000378:metabolism*; D011073:Polyamines / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D010455:Peptides; D010940:Plant Proteins; D011073:Polyamines; D029701:Saccharomyces cerevisiae Proteins; D009245:NADH Dehydrogenase