The Targeting of the atToc159 Preprotein Receptor to the Chloroplast Outer Membrane is Mediated by its GTPase Domain and is Regulated by GTP

The multimeric translocon at the outer envelope membrane of chloroplasts (Toc) initiates the recognition and import of nuclear-encoded preproteins into chloroplasts. Two Toc GTPases, Toc159 and Toc33/34, mediate preprotein recognition and regulate preprotein translocation. Although these two proteins account for the requirement of GTP hydrolysis for import, the functional significance of GTP binding and hydrolysis by either GTPase has not been defined. A recent study indicates that Toc159 is equally distributed between a soluble cytoplasmic form and a membrane-inserted form, raising the possibility that it might cycle between the cytoplasm and chloroplast as a soluble preprotein receptor. In the present study, we examined the mechanism of targeting and insertion of the Arabidopsis thaliana orthologue of Toc159, atToc159, to chloroplasts. Targeting of atToc159 to the outer envelope membrane is strictly dependent only on guanine nucleotides. Although GTP is not required for initial binding, the productive insertion and assembly of atToc159 into the Toc complex requires its intrinsic GTPase activity. Targeting is mediated by direct binding between the GTPase domain of atToc159 and the homologous GTPase domain of atToc33, the Arabidopsis Toc33/34 orthologue. Our findings demonstrate a role for the coordinate action of the Toc GTPases in assembly of the functional Toc complex at the chloroplast outer envelope membrane

Green aqueous surface modification of polypropylene for novel polymer nanocomposites

Polypropylene is one of the most widely used commercial commodity polymers; among many other applications, it is used for electronic and structural applications. Despite its commercial importance, the hydrophobic nature of polypropylene limits its successful application in some fields, in particular for the preparation of polymer nanocomposites. Here, a facile, plasma-assisted, biomimetic, environmentally friendly method was developed to enhance the interfacial interactions in polymer nanocomposites by modifying the surface of polypropylene. Plasma treated polypropylene was surface-modified with polydopamine (PDA) in an aqueous medium without employing other chemicals. The surface modification strategy used here was based on the easy self-polymerization and strong adhesion characteristics of dopamine (DA) under ambient laboratory conditions. The changes in surface characteristics of polypropylene were investigated using FTIR, TGA, and Raman spectroscopy. Subsequently, the surface modified polypropylene was used as the matrix to prepare SiO2-reinforced polymer nanocomposites. These nanocomposites demonstrated superior properties compared to nanocomposites prepared using pristine polypropylene. This simple, environmentally friendly, green method of modifying polypropylene indicated that polydopamine-functionalized polypropylene is a promising material for various high-performance applications

The targeting of the atToc159 preprotein receptor to the chloroplast outer membrane is mediated by its GTPase domain and is regulated by GTP

The multimeric translocon at the outer envelope membrane of chloroplasts (Toc) initiates the recognition and import of nuclear-encoded preproteins into chloroplasts. Two Toc GTPases, Toc159 and Toc33/34, mediate preprotein recognition and regulate preprotein translocation. Although these two proteins account for the requirement of GTP hydrolysis for import, the functional significance of GTP binding and hydrolysis by either GTPase has not been defined. A recent study indicates that Toc159 is equally distributed between a soluble cytoplasmic form and a membrane-inserted form, raising the possibility that it might cycle between the cytoplasm and chloroplast as a soluble preprotein receptor. In the present study, we examined the mechanism of targeting and insertion of the Arabidopsis thaliana orthologue of Toc159, atToc159, to chloroplasts. Targeting of atToc159 to the outer envelope membrane is strictly dependent only on guanine nucleotides. Although GTP is not required for initial binding, the productive insertion and assembly of atToc159 into the Toc complex requires its intrinsic GTPase activity. Targeting is mediated by direct binding between the GTPase domain of atToc159 and the homologous GTPase domain of atToc33, the Arabidopsis Toc33/34 orthologue. Our findings demonstrate a role for the coordinate action of the Toc GTPases in assembly of the functional Toc complex at the chloroplast outer envelope membrane

Essential Role of the G-Domain in Targeting of the Protein Import Receptor atToc159 to the Chloroplast Outer Membrane

Two homologous GTP-bindin proteins, atToc33 and atToc159, control access of cytosolic precursor proteins to the chloroplast, at Toc33 is a constitutive outer chloroplast membrane protein, whereas the precusor receptor atToc159 may be able to switch between a soluble and an integral membrane form. By transient expression of GFP fusion proteins, mutant analysis, and biochemical experimentation, we demonstrate that the GTP-binding domain regulates the targeting of cytosolic atToc159 to the chloroplast and mediates the switch between cytosolic and integral membrane forms. Mutant atToc159, unable to bind GTP, does not reinstate a green phenotype in an albino mutant (ppi2) lacking endogenous atToc159, remaining trapped in the cytosol. Thus, the function of atToc159 in chloroplast biogenesis is dependent on an intrinsic GTP-regulated swtich that controls localization of the receptor to the chloroplast envelope

How scent and nectar influence floral antagonists and mutualists

Abstract Many plants attract and reward pollinators with floral scents and nectar, respectively, but these traits can also incur fitness costs as they also attract herbivores. This dilemma, common to most flowering plants, could be solved by not producing nectar and/or scent, thereby cheating pollinators. Both nectar and scent are highly variable in native populations of coyote tobacco, Nicotiana attenuata, with some producing no nectar at all, uncorrelated with the tobacco's main floral attractant, benzylacetone. By silencing benzylacetone biosynthesis and nectar production in all combinations by RNAi, we experimentally uncouple these floral rewards/attractrants and measure their costs/benefits in the plant's native habitat and experimental tents. Both scent and nectar increase outcrossing rates for three, separately tested, pollinators and both traits increase oviposition by a hawkmoth herbivore, with nectar being more influential than scent. These results underscore that it makes little sense to study floral traits as if they only mediated pollination services

Substrate binding disrupts dimerization and induces nucleotide exchange of the chloroplast GTPase Toc33

GTPases act as molecular switches to control many cellular processes, including signalling, protein translation and targeting. Switch activity can be regulated by external effector proteins or intrinsic properties, such as dimerization. The recognition and translocation of pre-proteins into chloroplasts [via the TOC/TIC (translocator at the outer envelope membrane of chloroplasts/inner envelope membrane of chloroplasts)] is controlled by two homologous receptor GTPases, Toc33 and Toc159, whose reversible dimerization is proposed to regulate translocation of incoming proteins in a GTP-dependent manner. Toc33 is a homodimerizing GTPase. Functional analysis suggests that homodimerization is a key step in the translocation process, the molecular functions of which, as well as the elements regulating this event, are largely unknown. In the present study, we show that homodimerization reduces the rate of nucleotide exchange, which is consistent with the observed orientation of the monomers in the crystal structure. Pre-protein binding induces a dissociation of the Toc33 homodimer and results in the exchange of GDP for GTP. Thus homodimerization does not serve to activate the GTPase activity as discussed many times previously, but to control the nucleotide-loading state. We discuss this novel regulatory mode and its impact on the current models of protein import into the chloroplast

Natural variation in floral nectar proteins of two Nicotiana attenuata accessions

Background: Floral nectar (FN) contains not only energy-rich compounds to attract pollinators, but also defense chemicals and several proteins. However, proteomic analysis of FN has been hampered by the lack of publically available sequence information from nectar-producing plants. Here we used next-generation sequencing and advanced proteomics to profile FN proteins in the opportunistic outcrossing wild tobacco, Nicotiana attenuata. Results: We constructed a transcriptome database of N. attenuata and characterized its nectar proteome using LC-MS/MS. The FN proteins of N. attenuata included nectarins, sugar-cleaving enzymes (glucosidase, galactosidase, and xylosidase), RNases, pathogen-related proteins, and lipid transfer proteins. Natural variation in FN proteins of eleven N. attenuata accessions revealed a negative relationship between the accumulation of two abundant proteins, nectarin1b and nectarin5. In addition, microarray analysis of nectary tissues revealed that protein accumulation in FN is not simply correlated with the accumulation of transcripts encoding FN proteins and identified a group of genes that were specifically expressed in the nectary. Conclusions: Natural variation of identified FN proteins in the ecological model plant N. attenuata suggests that nectar chemistry may have a complex function in plant-pollinator-microbe interactions

Parental psychopathology, adult attachment and risk of 12-month suicidal behaviours

Background: The mechanisms by which parental psychopathology and vulnerability to suicide is transmitted to offspring is not well understood. parental psychopathology and behaviour may impact upon the normal emotional and psychological adjustment of their offspring in various ways. Research shows attachment insecurities may also be a key factor in the facilitation of suicidal behaviours. Objective: To examine adult attachment insecurities as a potential mediating pathway between parental psychopathology and 12- month suicidality. Method: The study utilized data from the National co-morbidity Survey- Replication (NCS-R, N=5692). Parental psychopathology was assessed using items from the Familial History of Psychiatric Disorders section of the NSC-R in conjunction with items designed to capture dimensions of attachment and suicidal behaviours. Results: Resultant analyses demonstrated specificity effects in that, parental psychopathology was associated with specific suicidal components through specific dimensions of attachment. Discussion: The results align with literature linking parental psychopathology to both attachment insecurities and risk of suicide. Crucially, this study bridges these research areas by presenting attachment insecurity as possible risk indicator and intervening factor between parental mental health and behaviour and specific indicators of suicide

Comparison of algorithms for incoming atmospheric long-wave radiation

While numerous algorithms exist for predicting incident atmospheric long-wave radiation under clear (Lclr) and cloudy skies, few comparisons have been published to assess the accuracy of the different algorithms. Virtually no comparisons have been made for both clear and cloudy skies across multiple sites. This study evaluates the accuracy of 13 algorithms for predicting incident long-wave radiation under clear skies, ten cloud correction algorithms, and four algorithms for all-sky conditions using data from 21 sites across North America and China. Data from five research sites were combined with publicly available data from nine sites in the AmeriFlux network for initial evaluation and optimization of cloud cover estimates; seven additional AmeriFlux sites were used as an independent test of the algorithms. Clear-sky algorithms that excelled in predicting Lclr were the Dilley, Prata, and Angström algorithms. Root mean square deviation (RMSD) between predicted and measured 30-minute or hourly Lclr averaged approximately 23 W m-2 for these three algorithms across all sites, while RMSD of daily estimates was as low as 14 W m-2. Cloud-correction algorithms of Kimball, Unsworth, and Crawford described the data best when combined with the Dilley clear-sky algorithm. Average RMSD across all sites for these three cloud corrections was approximately 24 to 25 W m -2 for 30-minute or hourly estimates and approximately 15 to 16 W m-2 for daily estimates. The Kimball and Unsworth cloud corrections require an estimate of cloud cover, while the Crawford algorithm corrects for cloud cover directly from measured solar radiation. Optimum limits in the clearness index, defined as the ratio of observed solar radiation to theoretical terrestrial solar radiation, for complete cloud cover and clear skies were suggested for the Kimball and Unsworth algorithms. Application of the optimized algorithms to seven independent sites yielded similar results. On the basis of the results, the recommended algorithms can be applied with reasonable accuracy for a wide range of climates, elevations, and latitudes. © 2009 by American Geophysical Union