Specialized functions of the PP2A subfamily II catalytic subunits PP2A-C3 and PP2A-C4 in the distribution of auxin fluxes and development in Arabidopsis.

Protein phosphorylation is a key molecular switch used to transmit information in biological signalling networks. The output of these signalling circuits is governed by the counteracting activities of protein kinases and phosphatases that determine the direction of the switch. Whereas many kinases have been functionally characterized, it has been difficult to ascribe precise cellular roles to plant phosphatases, which are encoded by enlarged gene families that may provide a high degree of genetic redundancy. In this work we have analysed the role in planta of catalytic subunits of protein phosphatase 2A (PP2A), a family encoded by five genes in Arabidopsis. Our results indicate that the two members of subfamily II, PP2A-C3 and PP2A-C4, have redundant functions in controlling embryo patterning and root development, processes that depend on auxin fluxes. Moreover, polarity of the auxin efflux carrier PIN1 and auxin distribution, determined with the DR5(pro) :GFP proxy, are affected by mutations in PP2A-C3 and PP2A-C4. Previous characterization of mutants in putative PP2A regulatory subunits had established a link between this class of phosphatases and PIN dephosphorylation and subcellular distribution. Building on those findings, the results presented here suggest that PP2A-C3 and PP2A-C4 catalyse this reaction and contribute critically to the establishment of auxin gradients for proper plant development.

Higher plant chloroplasts import the mRNA coding for the eucaryotic translation initiation factor 4E.

Plant chloroplasts probably originate from an endosymbiosis event between a photosynthetic bacteria and a eucaryotic cell. The proper functioning of this association requires a high level of integration between the chloroplastic genome and the plant cell genome. Many chloroplastic genes have been transferred to the nucleus of the host cell and the proteins coded by these genes are imported into the chloroplast. Chloroplastic activity also regulates the expression of these genes at the transcriptional and post-transcriptional levels. The importation of nucleic acids from the host cell into the chloroplast has never been observed. This work show that the mRNA coding for the eucaryotic translation factor 4E, an essential regulator of translation, enters the chloroplast in four different plant species, and is located in the stroma. Furthermore, the localization in the chloroplast of an heterologous GFP mRNA fused to the eIF4E RNA was also observed.

The Arabidopsis eukaryotic initiation factor (iso)4E is dispensable for plant growth but required for susceptibility to potyviruses.

An Arabidopsis thaliana line bearing a transposon insertion in the gene coding for the isozyme form of the plant-specific cap-binding protein, eukaryotic initiation factor (iso) 4E (eIF (iso) 4E), has been isolated. This mutant line completely lacks both eIF(iso)4E mRNA and protein, but was found to have a phenotype and fertility indistinguishable from wild-type plants under standard laboratory conditions. In contrast, the amount of the related eIF4E protein was found to increase in seedling extracts. Furthermore, polysome analysis shows that the mRNA encoding eIF4E was being translated at increased levels. Given the known interaction between cap-binding proteins and potyviral genome-linked proteins (VPg), this plant line was challenged with two potyviruses, Turnip mosaic virus (TuMV) and Lettuce mosaic virus (LMV) and was found resistant to both, but not to the Nepovirus, Tomato black ring virus (TBRV) and the Cucumovirus, Cucumber mosaic virus (CMV). Together with previous data showing that the VPg-eIF4E interaction is necessary for virus infectivity and upregulates genome amplification, this shows that the eIF4E proteins are specifically recruited for the replication cycle of potyviruses.

Effects of space environment on embryonic growth up to hatching of salamander eggs fertilized and developed during orbital flights.

In vertebrates, only few experiments have been performed in microgravity to study the embryonic development from fertilization. To date, these concern only amphibian and fish. We report here a study on the embryonic development of Pleurodeles waltl (urodele amphibian) eggs oviposited in microgravity. The experiment was performed twice on board the Mir space station and the data obtained included video recording and morphological, histological and immunocytological analyses. The data confirm that the microgravity conditions have effects during the embryonic period, particularly during cleavage and neurulation, inducing irregular segmentation and abnormal closure of the neural tube. Moreover, we observed several abnormalities hither to undescribed corresponding to cortical cytoplasm movements, a decrease of cell adhesion and a loss of cells. These abnormalities were temporary and subsequently reversible. The young larvae that hatched during the flight displayed normal morphology and swimming behavior after landing. The results obtained in the urodele Pleurodeles waltl are in accordance with those observed earlier in the anuran Xenopus laevis and in the fish Oryzias latipes.

[Inhibitory effects of homospecific non-histone chromatin proteins on morphogenesis of urodele amphibian embryos]. / Effets Inhibiteurs de Protéines Chromatiniennes non Histoniques Homospécifiques, sur la Morphogenèse d'Embryons d'Amphibiens Urodèles.

Non-histone chromosomal proteins (NHCP) were injected into the blastocoel of advanced blastulae fromPleurodeles waltlii which were previously punctured. NHCP extracted from the liver of the same amphibian species bring very strongly inhibitory effects on morphogenesis: gastrulation is prevented in 75% of cases, but the embryos can survive for 8 to 10 days without showing any necrotic cells, and cell divisions still occur sporadically. Intercellular adhesivity is strongly impaired in such embryos, in connection with inhibition of morphogenetic movements. Gastrulation is delayed and abnormal in other embryos, and neurulation is severely disturbed, the neural tube being unclosed or much reduced in size and the notochord and somites poorly differentiated. In contrast, NHCP extracted from other animal species (rat and chicken liver, rat ascites) have practically no effects onPleurodeles embryos. Species-specific inhibitory effects are thus demonstrated and compared with those which were previously studied on cultures of differentiating embryonic cells fromPleurodeles waltlii andAmbystoma mexicanum. The possible levels at which NHCP act are discussed, the changes in intercellular adhesivity being noted as more conspicuous in the present experiments.