Marc Hanikenne

Metal response of transgenic tomato plantsexpressing P1B-ATPase

Heterologous expression of HMA4 (P(1B) -ATPase) in plants is a useful strategy to engineer altered metal distribution in tissues for biofortification or phytoremediation purposes. This study contributes to understanding mechanisms underlying complex Zn-dependent phenotypes observed in transgenic plants and to better predict the consequences of transgene expression. Tomato was transformed with AhHMA4(p1) ::AhHMA4 from Arabidopsis halleri encoding the Zn export protein involved in xylem loading of Zn. Homozygous lines were tested for Zn tolerance, Zn and Fe concentrations in organs and in the apoplastic fluid, and for the expression of the transgene and tomato metal homeostasis endogenes. Expression of AhHMA4 facilitates root-to-shoot Zn translocation and induces Zn uptake in a Zn supply-dependent manner. Unexpectedly, it increases Zn excess-triggered Fe deficiency in leaves and transcriptional activation of Fe-uptake systems in roots. Moreover, AhHMA4 expression causes Zn overload of the apoplast, which may contribute to enhanced Zn sensitivity of transgenics and may lead to cell-wall remodeling. This study highlights that alteration of the apoplast/symplast Zn status through introduction of cellular Zn export activity via AhHMA4 may alter tomato metal homeostasis network, thus seems to be crucial in the generation of the phenotype of transgenic tomato.

A Single Ancient Origin for Prototypical Serine/Arginine-Rich Splicing Factors

Eukaryotic precursor mRNA splicing is a process involving a very complex RNA-protein edifice. Serine/arginine-rich (SR) proteins play essential roles in precursor mRNA constitutive and alternative splicing and have been suggested to be crucial in plant-specific forms of developmental regulation and environmental adaptation. Despite their functional importance, little is known about their origin and evolutionary history. SR splicing factors have a modular organization featuring at least one RNA recognition motif (RRM) domain and a carboxyl-terminal region enriched in serine/arginine dipeptides. To investigate the evolution of SR proteins, we infer phylogenies for more than 12,000 RRM domains representing more than 200 broadly sampled organisms. Our analyses reveal that the RRM domain is not restricted to eukaryotes and that all prototypical SR proteins share a single ancient origin, including the plant-specific SR45 protein. Based on these findings, we propose a scenario for their diversification into four natural families, each corresponding to a main SR architecture, and a dozen subfamilies, of which we profile both sequence conservation and composition. Finally, using operational criteria for computational discovery and classification, we catalog SR proteins in 20 model organisms, with a focus on green algae and land plants. Altogether, our study confirms the homogeneity and antiquity of SR splicing factors while establishing robust phylogenetic relationships between animal and plant proteins, which should enable functional analyses of lesser characterized SR family members, especially in green plants.

Chapter 10 – Transition Metal Nutrition: A Balance Between Deficiency and Toxicity

Publisher Summary This chapter describes the known components of the metal homeostasis network in Chlamydomonas, as well as the physiology and the molecular mechanisms of metal tolerance and metal deficiency responses. It delves into the metal tolerance mechanisms in Chlamydomonas and focuses on metal nutrition, deficiency, toxicity and tolerance. Unsuspected biological functions might be unveiled for these metals as more protein structures are elucidated and analytical techniques are improved. The discovery that Cd acts as the metal cofactor of a carbonic anhydrase isoform produced under Zn deficiency in the diatom Thalassiosira weissflogii suggests that metals previously considered to be non-essential and indeed toxic may occasionally act as a prosthetic group in biologically active metalloproteins. Physiological studies, genetic screens and functional analyses of candidate genes have advanced our understanding of the cellular adaptations to metal deficiency or overload in Chlamydomonas. The natural diversity of Chlamydomonas could be exploited to analyze the genetic variation associated with the metal homeostasis network within the species and genus.

Differential retention of transposable element-derived sequences in outcrossing Arabidopsis genomes

Transposable elements (TEs) have initially been viewed as pure genomic parasites but are now recognized as central genome architects and powerful agents of rapid adaptation. A proper evaluation of their evolutionary significance has been hampered by the paucity of short scale phylogenetic comparisons between closely related species. Here, we characterized the dynamics of TE accumulation at the micro-evolutionary scale by comparing two closely related plant species, Arabidopsis lyrata and A. halleri. Joint genome annotation in these two outcrossing species confirmed that both contain two distinct populations of TEs with either ‘recent’ or ‘old’ insertion histories. Identification of rare segregating insertions suggests that diverse TE families contribute to the ongoing dynamics of TE accumulation in the two species. TE fragments that have been maintained in both species, i.e. those that are orthologous, tend to be located on average closer to genes than those that are retained in one species only. Moreover, compared to non-orthologous TE insertions, those that are orthologous tend to produce fewer short interfering RNAs, are less heavily methylated when found within or adjacent to genes and these tend to have lower expression levels. These findings suggest that long-term retention of TE insertions reflects their frequent acquisition of adaptive roles and/or the deleterious effects of removing TE insertions when they are close to genes. Overall, our results indicate a rapid evolutionary dynamics of the TE landscape in these two outcrossing species, with an important input of a diverse set of new insertions with variable propensity to resist deletion.

di-Cysteine motifs in the C-terminus of plant HMA4 proteins confer nanomolar affinity for zinc and are essential for HMA4 function in vivo

Functional analysis of the HMA4 protein C-terminal extension reveals a similar and key function of high affinity zinc binding by di-Cys motifs in both Arabidopsis thaliana and A. halleri.