697 research outputs found
Modulation of plant growth in vivo and identification of kinase substrates using an analog-sensitive variant of CYCLIN-DEPENDENT KINASE A;1
BACKGROUND: Modulation of protein activity by phosphorylation through kinases and subsequent de-phosphorylation by phosphatases is one of the most prominent cellular control mechanisms. Thus, identification of kinase substrates is pivotal for the understanding of many – if not all – molecular biological processes. Equally, the possibility to deliberately tune kinase activity is of great value to analyze the biological process controlled by a particular kinase. RESULTS: Here we have applied a chemical genetic approach and generated an analog-sensitive version of CDKA;1, the central cell-cycle regulator in Arabidopsis and homolog of the yeast Cdc2/CDC28 kinases. This variant could largely rescue a cdka;1 mutant and is biochemically active, albeit less than the wild type. Applying bulky kinase inhibitors allowed the reduction of kinase activity in an organismic context in vivo and the modulation of plant growth. To isolate CDK substrates, we have adopted a two-dimensional differential gel electrophoresis strategy, and searched for proteins that showed mobility changes in fluorescently labeled extracts from plants expressing the analog-sensitive version of CDKA;1 with and without adding a bulky ATP variant. A pilot set of five proteins involved in a range of different processes could be confirmed in independent kinase assays to be phosphorylated by CDKA;1 approving the applicability of the here-developed method to identify substrates. CONCLUSION: The here presented generation of an analog-sensitive CDKA;1 version is functional and represent a novel tool to modulate kinase activity in vivo and identify kinase substrates. Our here performed pilot screen led to the identification of CDK targets that link cell proliferation control to sugar metabolism, proline proteolysis, and glucosinolate production providing a hint how cell proliferation and growth are integrated with plant development and physiology. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-016-0900-7) contains supplementary material, which is available to authorized users
Sampling Distributions of Random Electromagnetic Fields in Mesoscopic or Dynamical Systems
We derive the sampling probability density function (pdf) of an ideal
localized random electromagnetic field, its amplitude and intensity in an
electromagnetic environment that is quasi-statically time-varying statistically
homogeneous or static statistically inhomogeneous. The results allow for the
estimation of field statistics and confidence intervals when a single spatial
or temporal stochastic process produces randomization of the field. Results for
both coherent and incoherent detection techniques are derived, for Cartesian,
planar and full-vectorial fields. We show that the functional form of the
sampling pdf depends on whether the random variable is dimensioned (e.g., the
sampled electric field proper) or is expressed in dimensionless standardized or
normalized form (e.g., the sampled electric field divided by its sampled
standard deviation). For dimensioned quantities, the electric field, its
amplitude and intensity exhibit different types of
Bessel sampling pdfs, which differ significantly from the asymptotic
Gauss normal and ensemble pdfs when is relatively
small. By contrast, for the corresponding standardized quantities, Student ,
Fisher-Snedecor and root- sampling pdfs are obtained that exhibit
heavier tails than comparable Bessel pdfs. Statistical uncertainties
obtained from classical small-sample theory for dimensionless quantities are
shown to be overestimated compared to dimensioned quantities. Differences in
the sampling pdfs arising from de-normalization versus de-standardization are
obtained.Comment: 12 pages, 15 figures, accepted for publication in Phys. Rev. E, minor
typos correcte
SWITCH 1/DYAD is a WINGS APART-LIKE antagonist that maintains sister chromatid cohesion in meiosis
During cell division, Soronin antagonises WAPL to prevent premature loss of sister chromatid cohesion. Here the authors show that, despite a lack of sequence similarity, the Arabidopsis SWI1 protein functions as a novel Soronin-like WAPL antagonist, suggesting convergent evolution with animals
TOC1 clock protein phosphorylation controls complex formation with NF-YB/C to repress hypocotyl growth
Plant photoperiodic growth is coordinated by interactions between circadian clock and light signaling networks. How post‐translational modifications of clock proteins affect these interactions to mediate rhythmic growth remains unclear. Here, we identify five phosphorylation sites in the Arabidopsis core clock protein TIMING OF CAB EXPRESSION 1 (TOC1) which when mutated to alanine eliminate detectable phosphorylation. The TOC1 phospho‐mutant fails to fully rescue the clock, growth, and flowering phenotypes of the toc1 mutant. Further, the TOC1 phospho‐mutant shows advanced phase, a faster degradation rate, reduced interactions with PHYTOCHROME‐INTERACTING FACTOR 3 (PIF3) and HISTONE DEACETYLASE 15 (HDA15), and poor binding at pre‐dawn hypocotyl growth‐related genes (PHGs), leading to a net de‐repression of hypocotyl growth. NUCLEAR FACTOR Y subunits B and C (NF‐YB/C) stabilize TOC1 at target promoters, and this novel trimeric complex (NF‐TOC1) acts as a transcriptional co‐repressor with HDA15 to inhibit PIF‐mediated hypocotyl elongation. Collectively, we identify a molecular mechanism suggesting how phosphorylation of TOC1 alters its phase, stability, and physical interactions with co‐regulators to precisely phase PHG expression to control photoperiodic hypocotyl growth
Asymmetry Function of Interstellar Scintillations of Pulsars
A new method for separating intensity variations of a source's radio emission
having various physical natures is proposed. The method is based on a joint
analysis of the structure function of the intensity variations and the
asymmetry function, which is a generalization of the asymmetry coefficient and
characterizes the asymmetry of the distribution function of the intensity
fluctuations on various scales for the inhomogeneities in the diffractive
scintillation pattern. Relationships for the asymmetry function in the cases of
a logarithmic normal distribution of the intensity fluctuations and a normal
distribution of the field fluctuations are derived. Theoretical relationships
and observational data on interstellar scintillations of pulsars (refractive,
diffractive, and weak scintillations) are compared. Pulsar scintillations match
the behavior expected for a normal distribution of the field fluctuations
(diffractive scintillation) or logarithmic normal distribution of the intensity
fluctuations (refractive and weak scintillation). Analysis of the asymmetry
function is a good test for distinguishing scintillations against the
background of variations that have different origins
FLOWERING REPRESSOR AAA(+) ATPase 1 is a novel regulator of perennial flowering in Arabis alpina
Arabis alpina is a polycarpic perennial, in which PERPETUAL FLOWERING1 (PEP1) regulates flowering and perennial traits in a vernalization-dependent manner. Mutagenesis screens of the pep1 mutant established the role of other flowering time regulators in PEP1-parallel pathways. Here we characterized three allelic enhancers of pep1 (eop002, 085 and 091) which flower early. We mapped the causal mutations and complemented mutants with the identified gene. Using quantitative reverse transcriptase PCR and reporter lines, we determined the protein spatiotemporal expression patterns and localization within the cell. We also characterized its role in Arabidopsis thaliana using CRISPR and in A. alpina by introgressing mutant alleles into a wild-type background. These mutants carried lesions in an AAA(+) ATPase of unknown function, FLOWERING REPRESSOR AAA(+) ATPase 1 (AaFRAT1). AaFRAT1 was detected in the vasculature of young leaf primordia and the rib zone of flowering shoot apical meristems. At the subcellular level, AaFRAT1 was localized at the interphase between the endoplasmic reticulum and peroxisomes. Introgression lines carrying Aafrat1 alleles required less vernalization to flower and reduced number of vegetative axillary branches. By contrast, A. thaliana CRISPR lines showed weak flowering phenotypes. AaFRAT1 contributes to flowering time regulation and the perennial growth habit of A. alpina
Immunolocalization of dually phosphorylated MAPKs in dividing root meristem cells of Vicia faba, Pisum sativum, Lupinus luteus and Lycopersicon esculentum
Key message In plants, phosphorylated MAPKs display
constitutive nuclear localization; however, not all
studied plant species show co-localization of activated
MAPKs to mitotic microtubules.
Abstract The mitogen-activated protein kinase (MAPK)
signaling pathway is involved not only in the cellular
response to biotic and abiotic stress but also in the regulation
of cell cycle and plant development. The role of
MAPKs in the formation of a mitotic spindle has been
widely studied and the MAPK signaling pathway was
found to be indispensable for the unperturbed course of cell
division. Here we show cellular localization of activated
MAPKs (dually phosphorylated at their TXY motifs) in
both interphase and mitotic root meristem cells of Lupinus
luteus, Pisum sativum, Vicia faba (Fabaceae) and Lycopersicon esculentum (Solanaceae). Nuclear localization
of activated MAPKs has been found in all species. Colocalization
of these kinases to mitotic microtubules was
most evident in L. esculentum, while only about 50 % of
mitotic cells in the root meristems of P. sativum and V.
faba displayed activated MAPKs localized to microtubules
during mitosis. Unexpectedly, no evident immunofluorescence
signals at spindle microtubules and phragmoplast
were noted in L. luteus. Considering immunocytochemical
analyses and studies on the impact of FR180204 (an
inhibitor of animal ERK1/2) on mitotic cells, we hypothesize
that MAPKs may not play prominent role in the
regulation of microtubule dynamics in all plant species
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