Protein phosphatases PP2A, PP4 and PP6: mediators and regulators in development and responses to environmental cues.

The three closely related groups of serine/threonine protein phosphatases PP2A, PP4 and PP6 are conserved throughout eukaryotes. The catalytic subunits are present in trimeric and dimeric complexes with scaffolding and regulatory subunits that control activity and confer substrate specificity to the protein phosphatases. In Arabidopsis, three scaffolding (A subunits) and 17 regulatory (B subunits) proteins form complexes with five PP2A catalytic subunits giving up to 255 possible combinations. Three SAP-domain proteins act as regulatory subunits of PP6. Based on sequence similarities with proteins in yeast and mammals, two putative PP4 regulatory subunits are recognized in Arabidopsis. Recent breakthroughs have been made concerning the functions of some of the PP2A and PP6 regulatory subunits, for example the FASS/TON2 in regulation of the cellular skeleton, B' subunits in brassinosteroid signalling and SAL proteins in regulation of auxin transport. Reverse genetics is starting to reveal also many more physiological functions of other subunits. A system with key regulatory proteins (TAP46, TIP41, PTPA, LCMT1, PME-1) is present in all eukaryotes to stabilize, activate and inactivate the catalytic subunits. In this review, we present the status of knowledge concerning physiological functions of PP2A, PP4 and PP6 in Arabidopsis, and relate these to yeast and mammals.

PP2A activates brassinosteroid-responsive gene expression and plant growth by dephosphorylating BZR1.

When brassinosteroid levels are low, the GSK3-like kinase BIN2 phosphorylates and inactivates the BZR1 transcription factor to inhibit growth in plants. Brassinosteroid promotes growth by inducing dephosphorylation of BZR1, but the phosphatase that dephosphorylates BZR1 has remained unknown. Here, using tandem affinity purification, we identified protein phosphatase 2A (PP2A) as a BZR1-interacting protein. Genetic analyses demonstrated a positive role for PP2A in brassinosteroid signalling and BZR1 dephosphorylation. Members of the B' regulatory subunits of PP2A directly interact with BZR1's putative PEST domain containing the site of the bzr1-1D mutation. Interaction with and dephosphorylation by PP2A are enhanced by the bzr1-1D mutation, reduced by two intragenic bzr1-1D suppressor mutations, and abolished by deletion of the PEST domain. This study reveals a crucial function for PP2A in dephosphorylating and activating BZR1 and completes the set of core components of the brassinosteroid-signalling cascade from cell surface receptor kinase to gene regulation in the nucleus.

The bZIP transcription factors HY5 and HYH are positive regulators of the main nitrate reductase gene in Arabidopsis leaves, NIA2, but negative regulators of the nitrate uptake gene NRT1.1.

Previously we have shown that nitrate reductase (NR) activity was impaired in Arabidopsis seedlings and rosette stage leaves when the basic leucine zipper transcription factors HY5 and HYH were absent. In the present work, we investigated the influence of hy5 and hyh null mutations on the expression of the NR encoding genes NIA1 and NIA2, as well as genes involved in nitrate uptake and further assimilation of nitrite. Only NIA2, and not NIA1, transcript levels were positively influenced by the presence of HY5 and HYH. In the hy5 hyh mutant, enhancement of NIA2 expression by light was impaired in both seedlings and rosette stage plants. Induction of NIA2 transcription by nitrate was not influenced by HY5 or HYH. Although the peak NIA2 transcript level was much higher in wild type in comparison with the hy5 hyh mutant plant, diurnal variations of NIA2 expression were also observed in the mutant. A dual affinity nitrate transporter gene, NRT1.1, was expressed at a higher level in hy5 hyh than in the wild type, indicating that HY5 and HYH are repressors of NRT1.1. In conclusion, HY5 and HYH were activators of NIA2, but inhibitors of NRT1.1 when tested across various light treatments and tissue types.