The Arabidopsis epitranscriptome

The most prevalent internal modification of plant messenger RNAs, N6-methyladenosine (m6A), was first discovered in the 1970s, then largely forgotten. However, the impact of modifications to eukaryote mRNA, collectively known as the epitranscriptome, has recently attracted renewed attention. mRNA methylation is required for normal Arabidopsis development and the first methylation maps reveal that thousands of Arabidopsis mRNAs are methylated. Arabidopsis is likely to be a model of wide utility in understanding the biological impacts of the epitranscriptome. We review recent progress and look ahead with questions awaiting answers to reveal an entire layer of gene regulation that has until recently been overlooked

Improved plant transformation vectors for fluorescent protein tagging

Fluorescent protein labelling technologies enable dynamic protein actions to be imaged in living cells and can also be used in conjunction with other methods such as Forster resonance energy transfer and biomolecular fluorescence complementation. In this report, we describe the generation of a series of 23 novel GATEWAY-compatible vectors based on pGreenII and pDH51 backbones with the latest fluorescent protein tags (Cerulean, EGFP and Venus) and the choice of three in planta selection markers. These vectors can be obtained from the Nottingham Arabidopsis Stock Centre (N9819-N9846) and should be a powerful tool box for transgenic research in plants

Methylation of chloroplast DNA does not affect viability and maternal inheritance in tobacco and may provide a strategy towards transgene containment

We report the integration of a type II restriction-methylase, mFokI, into the tobacco chloroplast genome and we demonstrate that the introduced enzyme effectively directs the methylation of its target sequence in vivo and does not affect maternal inheritance. We further report the transformation of tobacco with an E. coli dcm methylase targeted to plastids and we demonstrate efficient cytosine methylation of the plastid genome. Both adenosine methylation of FokI sites and cytosine methylation of dcm sites appeared phenotypically neutral. The ability to tolerate such plastid genome methylation is a pre-requisite for a proposed plant transgene containment system. In such a system, a chloroplast located, maternally inherited restriction methylase would provide protection from a nuclear-encoded, plastid targeted restriction endonuclease. As plastids are not paternally inherited in most crop species, pollen from such plants would carry the endonuclease transgene but not the corresponding methylase; the consequence of this should be containment of all nuclear transgenes, as pollination will only be viable in crosses to the appropriate transplastomic maternal background

Yeast m6 A methylated mRNAs are enriched on translating ribosomes during meiosis, and under rapamycin treatment

Interest in mRNA methylation has exploded in recent years. The sudden interest in a 40 year old discovery was due in part to the finding of FTO’s (Fat Mass Obesity) N6-methyladenosine (m6 A) deaminase activity, thus suggesting a link between obesity-associated diseases and the presence of m6 A in mRNA. Another catalyst of the sudden rise in mRNA methylation research was the release of mRNA methylomes for human, mouse and Saccharomyces cerevisiae. However, the molecular function, or functions of this mRNA ‘epimark’ remain to be discovered. There is supportive evidence that m6 A could be a mark for mRNA degradation due to its binding to YTH domain proteins, and consequently being chaperoned to P bodies. Nonetheless, only a subpopulation of the methylome was found binding to YTHDF2 in HeLa cells.The model organism Saccharomyces cerevisiae, has only one YTH domain protein (Pho92, Mrb1), which targets PHO4 transcripts for degradation under phosphate starvation. However, mRNA methylation is only found under meiosis inducing conditions, and PHO4 transcripts are apparently non-methylated. In this paper we set out to investigate if m6 A could function alternatively to being a degradation mark in S. cerevisiae; we also sought to test whether it can be induced under non-standard sporulation conditions. We find a positive association between the presence of m6 A and message translatability. We also find m6 A induction following prolonged rapamycin treatment

Low CO/CO₂ ratios of comet 67P measured at the Abydos landing site by the <i>Ptolemy</i> mass spectrometer

Comets are generally considered to contain the best-preserved material from the beginning of our planetary system, although the mechanism of their formation and subsequent evolution are still poorly understood. Here we report the direct in situ measurement of H2O, CO, and CO2 by the Ptolemy mass spectrometer onboard the Philae lander, part of the European Space Agency’s Rosetta mission, at the Abydos site of the Jupiter-family comet 67P/Churyumov-Gerasimenko. A CO/CO2 ratio of around 0.07 ± 0.04 is found at the surface of the comet, a value substantially lower than the one measured by ROSINA in the coma. Such a major difference is a potential indication of heterogeneity of the nucleus and not of changes in the CO/CO2 ratio of the coma with radial distance

Rotation-stimulated structures in the CN and C3 comae of comet 103P/Hartley 2 around the EPOXI encounter

In late 2010 a Jupiter Family comet 103P/Hartley 2 was a subject of an intensive world-wide investigation. On UT October 20.7 the comet approached the Earth within only 0.12 AU, and on UT November 4.6 it was visited by NASA's EPOXI spacecraft. We joined this international effort and organized an observing campaign. The images of the comet were obtained through narrowband filters using the 2-m telescope of the Rozhen National Astronomical Observatory. They were taken during 4 nights around the moment of the EPOXI encounter. Image processing methods and periodicity analysis techniques were used to reveal transient coma structures and investigate their repeatability and kinematics. We observe shells, arc-, jet- and spiral-like patterns, very similar for the CN and C3 comae. The CN features expanded outwards with the sky-plane projected velocities between 0.1 to 0.3 km/s. A corkscrew structure, observed on November 6, evolved with a much higher velocity of 0.66 km/s. Photometry of the inner coma of CN shows variability with a period of 18.32+/-0.30 h (valid for the middle moment of our run, UT 2010 Nov. 5.0835), which we attribute to the nucleus rotation. This result is fully consistent with independent determinations around the same time by other teams. The pattern of repeatability is, however, not perfect, which is understendable given the suggested excitation of the rotation state, and the variability detected in CN correlates well with the cyclic changes in HCN, but only in the active phases. The revealed coma structures, along with the snapshot of the nucleus orientation obtained by EPOXI, let us estimate the spin axis orientation. We obtained RA=122 deg, Dec=+16 deg (epoch J2000.0), neglecting at this point the rotational excitation.Comment: 9 pages, 10 figures, submitted to Astron. Astrophy

Two zinc finger proteins with functions in m6A writing interact with HAKAI

The methyltransferase complex (m6A writer), which catalyzes the deposition of N6-methyladenosine (m6A) in mRNAs, is highly conserved across most eukaryotic organisms, but its components and interactions between them are still far from fully understood. Here, using in vivo interaction proteomics, two HAKAI-interacting zinc finger proteins, HIZ1 and HIZ2, are discovered as components of the Arabidopsis m6A writer complex. HAKAI is required for the interaction between HIZ1 and MTA (mRNA adenosine methylase A). Whilst HIZ1 knockout plants have normal levels of m6A, plants in which it is overexpressed show reduced methylation and decreased lateral root formation. Mutant plants lacking HIZ2 are viable but have an 85% reduction in m6A abundance and show severe developmental defects. Our findings suggest that HIZ2 is likely the plant equivalent of ZC3H13 (Flacc) of the metazoan m6A-METTL Associated Complex

Niobium based intermetallics as a source of high-current/high-magnetic field superconductors

The article is focused on low temperature intermetallic A15 superconducting wires development for Nuclear Magnetic Resonance, NMR, and Nuclear Magnetic Imaging, MRI, magnets and also on cryogen-free magnets. There are many other applications which would benefit from new development such as future Large Hadron Collider to be built from A15 intermetallic conductors. This paper highlights the current status of development of the niobium based intermetallics with special attention to Nb 3 (Al 1-x, Ge x). Discussion is focused on the materials science aspects of conductor manufacture, such as b-phase (A15) formation, with particular emphasis on the maximisation of the superconducting parameters, such as critical current density, Jc, critical temperature, Tc, and upper critical field, Hc2 . Many successful manufacturing techniques of the potential niobium-aluminide intermetallic superconducting conductors, such as solid-state processing, liquid-solid processing, rapid heating/cooling processes, are described, compared and assessed. Special emphasis has been laid on conditions under which the Jc (B) peak effect occurs in some of the Nb3(Al,Ge) wires. A novel electrodeoxidizing method developed in Cambridge whereby the alloys and intermetallics are produced cheaply making all superconducting electromagnetic devices, using low cost LTCs, more cost effective is presented.This new technique has potential to revolutionise the existing superconducting industry enabling reduction of cost orders of magnitude.Comment: Paper presented at EUCAS'01 conference, Copenhagen, 26-30 August 200

Exogenous N-acyl-homoserine lactones enhance the expression of flagella of Pseudomonas syringae and activate defence responses in plants

In order to cope with pathogens, plants have evolved sophisticated mechanisms to sense pathogenic attacks and to induce defence responses. The N-acyl-homoserine lactone (AHL)-mediated quorum sensing in bacteria regulates diverse physiological processes, including those involved in pathogenicity. In this work, we study the interactions between AHL-producing transgenic tobacco plants and Pseudomonas syringae pv. tabaci 11528 (P. syringae 11528). Both a reduced incidence of disease and decrease in the growth of P. syringae 11528 were observed in AHL-producing plants compared with wild-type plants. The present data indicate that plant-produced AHLs enhance disease resistance against this pathogen. Subsequent RNA-sequencing analysis showed that the exogenous addition of AHLs upregulated the expression of P. syringae 11528 genes for flagella production. Expression levels of plant defence genes in AHL producing and wild-type plants were determined by quantitative real-time polymerase chain reaction. These data showed that plant-produced AHLs activated a wide spectrum of defence responses in plants following inoculation, including the oxidative burst, hypersensitive response, cell wall strengthening, and the production of certain metabolites. These results demonstrate that exogenous AHLs alter the gene expression patterns of pathogens, and plant-produced AHLs either directly or indirectly enhance plant local immunity during the early stage of plant infection