Plant cells respond to different biotic and abiotic stresses by producing various uncommon phospholipids that are believed to play key roles in cell signalling. We can predict how they work because animal and yeast proteins have been shown to have specific lipid-binding domains, which act as docking sites. When such proteins are recruited to the membrane locations where these phospholipids are synthesized, the phospholipids activate them directly, by inducing a conformational change, or indirectly, by juxtaposing them with an activator protein. The same lipid-binding domains are present in Arabidopsis proteins. We believe that they represent an untapped well of information about plant lipid signalling.

The anaphase-promoting complex or cyclosome (APC/C) is a cell-cycle-regulated ubiquitin-protein ligase that has been extensively studied in both fungal and animal cells. Many APC/C protein targets have been identified, and their sequential degradation during the cell cycle is essential for chromatid separation and mitotic exit. APC/C-dependent ubiquitylation of proteins not involved in cell-cycle progression has also been documented in animal cells. By contrast, the plant APC/C's structure and functions remained unexplored until recently. Here, we discuss recent developments in this field and explore the Arabidopsis genome sequence to identify plant APC/C components. Details of the APC/C ubiquitylation pathway in Arabidopsis are also available on a website that will be regularly updated.

Lipid-derived signals are central to regulating a multitude of cellular processes but, in plants, little is known of the downstream signalling pathways. The Arabidopsis 3-phosphoinositide-dependent protein kinase (PDK1) could couple lipid signals to the activation of several protein kinases of the so-called AGC kinase family. The Arabidopsis AGC kinases contain sequence motives required for the docking of PDK1 and phosphorylation of their activation loop in the kinase catalytic domain. It is becoming evident that specific members of the AGC kinases are implicated in key growth signalling pathways. For example, Arabidopsis p70S6K might be a nodal point able to integrate hormonal and developmental signals with nutritional inputs, together with the Arabidopsis Target of Rapamycin (TOR) protein.

Mitogen-activated protein kinases (MAPKs) link information transfer from external stimuli-activated sensors to cellular responses. The completed Arabidopsis genome sequence revealed an extraordinary complexity in MAPK-signalling components in plants, having an overall of 123 MAPK-, MAPKK-, MAPKKK- and MAPKKKK-related genes. Arabidopsis provides a framework for a unified nomenclature and the assembly and function of MAPK-signalling pathways. Strategies and tools are evolving to connect MAPK pathways and to determine their function. As a result, MAPK signalling modules emerged, one of which appears to antagonistically regulate stress- and growth-responses and another that regulates cytokinesis. This review focusses on recent advances in exploring the phylogeny and function of MAPK signalling in plants.

Arabidopsis Snf1‐related protein kinases (SnRKs) are implicated in pleiotropic regulation of metabolic, hormonal and stress responses through their interaction with the kinase inhibitor PRL1 WD‐protein. Here we show that SKP1/ASK1, a conserved SCF (Skp1‐cullin‐F‐box) ubiquitin ligase subunit, which suppresses the skp1‐4 mitotic defect in yeast, interacts with the PRL1‐binding C‐terminal domains of SnRKs. The same SnRK domains recruit an SKP1/ASK1‐binding proteasomal protein, α4/PAD1, which enhances the formation of a trimeric SnRK complex with SKP1/ASK1 in vitro. By contrast, PRL1 reduces the interaction of SKP1/ASK1 with SnRKs. SKP1/ASK1 is co‐immunoprecipitated with a cullin SCF subunit (AtCUL1) and an SnRK kinase, but not with PRL1 from Arabidopsis cell extracts. SKP1/ASK1, cullin and proteasomal α‐subunits show nuclear co‐localization in differentiated Arabidopsis cells, and are observed in association with mitotic spindles and phragmoplasts during cell division. Detection of SnRK in purified 26S proteasomes and co‐purification of epitope‐ tagged SKP1/ASK1 with SnRK, cullin and proteasomal α‐subunits indicate that the observed protein interactions between SnRK, SKP1/ASK1 and α4/PAD1 are involved in proteasomal binding of an SCF ubiquitin ligase in Arabidopsis.

Mutation of the PRL1 gene, encoding a regulatory WD protein, results in glucose hypersensitivity and derepression of glucose-regulated genes in Arabidopsis. The yeast SNF1 protein kinase, a key regulator of glucose signaling, and Arabidopsis SNF1 homologs AKIN10 and AKIN11, which can complement the Deltasnf1 mutation, were found to interact with an N-terminal domain of the PRL1 protein in the two-hybrid system and in vitro. AKIN10 and AKIN11 suppress the yeast Deltasnf4 mutation and interact with the SNF4p-activating subunit of SNF1. PRL1 and SNF4 bind independently to adjacent C-terminal domains of AKIN10 and AKIN11, and these protein interactions are negatively regulated by glucose in yeast. AKIN10 and AKIN11, purified in fusion with glutathione S-transferase, undergo autophosphorylation and phosphorylate a peptide of sucrose phosphate synthase in vitro. The sucrose phosphate synthase-peptide kinase activity of AKIN complexes detected by immunoprecipitation is stimulated by sucrose in light-grown Arabidopsis plants. In comparison with wild type, the activation level of AKIN immunocomplexes is higher in the prl1 mutant, suggesting that PRL1 is a negative regulator of Arabidopsis SNF1 homologs. This conclusion is supported by the observation that PRL1 is an inhibitor of AKIN10 and AKIN11 in vitro.

The prl1 mutation localized by T-DNA tagging on Arabidopsis chromosome 4-44 confers hypersensitivity to glucose and sucrose. The prl1 mutation results in transcriptional derepression of glucose responsive genes defining a novel suppressor function in glucose signaling. The prl1 mutation also augments the sensitivity of plants to growth hormones including cytokinin, ethylene, abscisic acid, and auxin; stimulates the accumulation of sugars and starch in leaves; and inhibits root elongation. PRL1 encodes a regulatory WD protein that interacts with ATHKAP2, an alpha-importin nuclear import receptor, and is imported into the nucleus in Arabidopsis. Potential functional conservation of PRL1 homologs found in other eukaryotes is indicated by nuclear localization of PRL1 in monkey COS-1 cells and selective interaction of PRL1 with a nuclear protein kinase C-beta II isoenzyme involved in human insulin signaling.

Proline is a common compatible osmolyte in higher plants. Proline accumulation in response to water stress and salinity is preceded by a rapid increase of the mRNA level of Delta 1-pyrroline-5-carboxylate synthase (P5CS) controlling the rate-limiting step of glutamate-derived proline biosynthesis. P5CS is encoded by two differentially regulated genes in Arabidopsis. Gene AtP5CS1 mapped to chromosome 2-78.5 is expressed in most plant organs, but silent in dividing cells. Gene AtP5CS2 located close to marker m457 on chromosome 3-101.3, and is responsible for the synthesis of abundant P5CS mRNA in dividing cells. Accumulation of AtP5CS transcripts is regulated in a tissue specific manner and inducible by drought, salinity, ABA, and to lesser extent by auxin. Induction of AtP5CS1 mRNA accumulation in salt-treated seedlings involves an immediate early transcriptional response regulated by ABA signaling. Inhibition of protein synthesis by cycloheximide affects the induction of AtP5CS mRNA accumulation. Mutations aba1, abi1 and axr2, affecting ABA synthesis and perception in Arabidopsis, reduce the accumulation of both AtP5CS mRNAs during salt-stress, whereas ABA-signaling functions defined by the abi2 and abi3 mutations have no effect on salt-induction of the AtP5CS genes. Promoter regions of the AtP5CS genes has been cloned and the sequence of 1 kb. fragments has been determined. Sequence analysis of the promoter regions of the AtP5CS genes revealed the presence of putative conserved cis-acting elements, including a G-box. cDNA of AtP5CS1 has been placed under the control of different plant promoter sequences in binary expression vectors. Genetic transformation of tobacco and alfalfa resulted in the regeneration of transgenic plants containing increased internal proline content. Studies on the effect of proline accumulation on salt and drought tolerance is in progress.

A T-DNA tagged Arabidopsis locus, A37, identified by a promoter-trap aph(3′)II reporter gene fusion expressed in calli and roots, encodes an ortholog of evolutionarily conserved SNZ growth arrest response proteins. Gene A37 is located on chromosome 3–35, lacks introns, and shares considerable sequence identity with HEVER1 from rubber tree, SLEXORFA-1 from Stellaria longipes, SNZ1 from yeast, and SNZ-homologs from bacteria and archaebacteria. Southern DNA hybridization and physical mapping data show that A37 is a single copy gene, but sequence similarity to expressed sequence tags (ESTs) suggests that at least two other SNZ-homologs are present in Arabidopsis. The A37 gene is abundantly expressed in cultured callus tissues and at lower levels in leaves, stems and roots. In the promoter-trap locus a37, the T-DNA-linked aph(3′)II reporter gene is transcribed oppositely to the A37 gene by a cryptic promoter located 0.52 kb upstream of the A37 coding region. Promoter deletion studies with uidA-reporter gene constructs show that the cryptic promoter consists of regulatory sequences located in both promoter and transcribed regions of the A37 gene that activate transcription only in roots. The a37 promoter trap is thus controlled by transcriptional regulatory sequences that function as an active promoter only in linkage with a promoterless reporter gene introduced artificially into the Arabidopsis genome by a T-DNA tag.

Proline is a common compatible osmolyte in higher plants. Proline accumulation in response to water stress and salinity is preceded by a rapid increase of the mRNA level of delta 1-pyrroline-5-carboxylate synthase (P5CS) controlling the rate-limiting step of glutamate-derived proline biosynthesis. P5CS is encoded by two differentially regulated genes in Arabidopsis. Gene AtP5CS1 mapped to chromosome 2-78.5 is expressed in most plant organs, but silent in dividing cells. Gene AtP5CS2 located close to marker m457 on chromosome 3-101.3 contributes 20-40% of total P5CS mRNA in plant tissues, but is solely responsible for the synthesis of abundant P5CS mRNA in rapidly dividing cell cultures. Accumulation of AtP5CS transcripts is regulated in a tissue specific manner and inducible by drought, salinity, ABA, and to a lesser extent by auxin. Induction of AtP5CS1 mRNA accumulation in salt-treated seedlings involves an immediate early transcriptional response regulated by ABA signalling that is not inhibited by cycloheximide, but abolished by the deficiency of ABA biosynthesis in the aba1 Arabidopsis mutant. However, inhibition of protein synthesis by cycloheximide prevents the induction of AtP5CS2 mRNA accumulation, and blocks further increase of AtP5CS1 mRNA levels during the second, slow phase of salt-induction. Mutations abi1 and axr2, affecting ABA-perception in Arabidopsis, reduce the accumulation of both AtP5CS mRNAs during salt-stress, whereas ABA-signalling functions defined by the abi2 and abi3 mutations have no effect on salt-induction of the AtP5CS genes.