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bioRxiv · 10.64898/2026.09.03.749138

The PP2A phosphatase associates with the Arabidopsis TRAPPII tethering complex and dephosphorylates a TRAPPII-derived phosphopeptide in vitro

Strohmayr, A.·Steiner, A.·Wiese, C.·Abele, M.·Facher, E.·Belcram, K.·Altmann, M.·Falter-Braun, P.·Ludwig, C.·Pastuglia, M.·Bouchez, D.·Assaad, F. F.
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Abstract

The transport protein particle II (TRAPPII) complex is a conserved regulator of post-Golgi membrane trafficking. In Arabidopsis, phosphorylation of the TRAPPII-specific subunit TRS120 by SHAGGY-like kinases modulates adaptive growth responses, but the phosphatases that reverse this phosphorylation remain unknown. Here, proteomic analyses identified subunits of Protein Phosphatase 2A (PP2A) in the TRAPPII interactome. PP2A subunits physically associated with TRAPPII, and double-mutant analyses revealed genetic interactions between PP2A and TRAPPII. Loss of TRAPPII function reduced the relative membrane association of PP2A scaffolding subunits. We established an in vitro assay for Arabidopsis PP2A holoenzyme activity using complexes transiently co-expressed and affinity-purified from Nicotiana benthamiana. Structural modelling and interface analysis predicted binding of a phosphorylated peptide encompassing a TRS120 phosphosite cluster at the PP2A catalytic interface, while biochemical assays showed that a PP2A holoenzyme containing the B2 regulatory subunit dephosphorylated this peptide. Together, these findings identify a B2-containing PP2A holoenzyme as a candidate phosphatase for TRS120 and support a model in which antagonistic SHAGGY-like kinase and PP2A activities couple signalling to membrane trafficking during plant development and environmental adaptation.

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Strohmayr, A., Steiner, A., Wiese, C., Abele, M., Facher, E., Belcram, K., Altmann, M., Falter-Braun, P., Ludwig, C., Pastuglia, M., Bouchez, D., Assaad, F. F.. 2026-09-07. The PP2A phosphatase associates with the Arabidopsis TRAPPII tethering complex and dephosphorylates a TRAPPII-derived phosphopeptide in vitro. https://doi.org/10.64898/2026.09.03.749138

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