56 research outputs found
Diverse Physiological Functions of FAB1 and Phosphatidylinositol 3,5-Bisphosphate in Plants
Biological membranes are predominantly composed of structural glycerophospholipids such as phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. Of the membrane glycerophospholipids, phosphatidylinositol (PtdIns) and its phosphorylated derivatives (phosphoinositides) constitute a minor fraction yet exert a wide variety of regulatory functions in eukaryotic cells. Phosphoinositides include PtdIns, three PtdIns monophosphates, three PtdIns bisphosphates, and one PtdIns triphosphate, in which the hydroxy groups of the inositol head group of PtdIns are phosphorylated by specific lipid kinases. Of all the phosphoinositides in eukaryotic cells, phosphatidylinositol 3,5-bisphosphate [PtdIns(3,5)P2] constitutes the smallest fraction, yet it is a crucial lipid in animal and yeast membrane trafficking systems. Here, we review the recent findings on the physiological functions of PtdIns(3,5)P2 and its enzyme—formation of aploid and binucleate cells (FAB1)—along with the regulatory proteins of FAB1 and the downstream effector proteins of PtdIns(3,5)P2 in Arabidopsis
Metal Ion Homeostasis Mediated by NRAMP Transporters in Plant Cells - Focused on Increased Resistance to Iron and Cadmium Ion
Atomic force microscopy sees nucleosome positioning and histone H1-induced compaction in reconstituted chromatin
AbstractWe addressed the question of how nuclear histones and DNA interact and form a nucleosome structure by applying atomic force microscopy to an in vitro reconstituted chromatin system. The molecular images obtained by atomic force microscopy demonstrated that oligonucleosomes reconstituted with purified core histones and DNA yielded a ‘beads on a string’ structure with each nucleosome trapping 158±27 bp DNA. When dinucleosomes were assembled on a DNA fragment containing two tandem repeats of the positioning sequence of the Xenopus 5S RNA gene, two nucleosomes were located around each positioning sequence. The spacing of the nucleosomes fluctuated in the absence of salt and the nucleosomes were stabilized around the range of the positioning signals in the presence of 50 mM NaCl. An addition of histone H1 to the system resulted in a tight compaction of the dinucleosomal structure
Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format
© 2014 Fujikawa et al. Background: The rice interactome, in which a network of protein-protein interactions has been elucidated in rice, is a useful resource to identify functional modules of rice signal transduction pathways. Protein-protein interactions occur in cells in two ways, constitutive and regulative. While a yeast-based high-throughput method has been widely used to identify the constitutive interactions, a method to detect the regulated interactions is rarely developed for a large-scale analysis. Results: A split luciferase complementation assay was applied to detect the regulated interactions in rice. A transformation method of rice protoplasts in a 96-well plate was first established for a large-scale analysis. In addition, an antibody that specifically recognizes a carboxyl-terminal fragment of Renilla luciferase was newly developed. A pair of antibodies that recognize amino- and carboxyl- terminal fragments of Renilla luciferase, respectively, was then used to monitor quality and quantity of interacting recombinant-proteins accumulated in the cells. For a proof-of-concept, the method was applied to detect the gibberellin-dependent interaction between GIBBERELLIN INSENSITIVE DWARF1 and SLENDER RICE 1. Conclusions: A method to detect regulated protein-protein interactions was developed towards establishment of the rice interactome
Heavy Element Production in Inhomogeneous Big Bang Nucleosynthesis
We present a new astrophysical site of the big bang nucleosynthesis (BBN)
that are very peculiar compared with the standard BBN. Some models of the
baryogenesis suggest that very high baryon density regions were formed in the
early universe. On the other hand, recent observations suggest that heavy
elements already exist in high red-shifts and the origin of these elements
become a big puzzle. Motivated by these, we investigate BBN in very high baryon
density regions. BBN proceeds in proton-rich environment, which is known to be
like the p-process. However, by taking very heavy nuclei into account, we find
that BBN proceeds through both the p-process and the r-process simultaneously.
P-nuclei such as 92Mo, 94Mo, 96Ru, 98Ru whose origin is not well known are also
synthesized.Comment: 6 pages, 7 figure
Ultra-stable performance of an underground-based laser interferometer observatory for gravitational waves
In order to detect the rare astrophysical events that generate gravitational
wave (GW) radiation, sufficient stability is required for GW antennas to allow
long-term observation. In practice, seismic excitation is one of the most
common disturbances effecting stable operation of suspended-mirror laser
interferometers. A straightforward means to allow more stable operation is
therefore to locate the antenna, the ``observatory'', at a ``quiet'' site. A
laser interferometer gravitational wave antenna with a baseline length of 20m
(LISM) was developed at a site 1000m underground, near Kamioka, Japan. This
project was a unique demonstration of a prototype laser interferometer for
gravitational wave observation located underground. The extremely stable
environment is the prime motivation for going underground. In this paper, the
demonstrated ultra-stable operation of the interferometer and a well-maintained
antenna sensitivity are reported.Comment: 8 pages, to appear on PR
Coincidence analysis to search for inspiraling compact binaries using TAMA300 and LISM data
Japanese laser interferometric gravitational wave detectors, TAMA300 and
LISM, performed a coincident observation during 2001. We perform a coincidence
analysis to search for inspiraling compact binaries. The length of data used
for the coincidence analysis is 275 hours when both TAMA300 and LISM detectors
are operated simultaneously. TAMA300 and LISM data are analyzed by matched
filtering, and candidates for gravitational wave events are obtained. If there
is a true gravitational wave signal, it should appear in both data of detectors
with consistent waveforms characterized by masses of stars, amplitude of the
signal, the coalescence time and so on. We introduce a set of coincidence
conditions of the parameters, and search for coincident events. This procedure
reduces the number of fake events considerably, by a factor
compared with the number of fake events in single detector analysis. We find
that the number of events after imposing the coincidence conditions is
consistent with the number of accidental coincidences produced purely by noise.
We thus find no evidence of gravitational wave signals. We obtain an upper
limit of 0.046 /hours (CL ) to the Galactic event rate within 1kpc from
the Earth. The method used in this paper can be applied straightforwardly to
the case of coincidence observations with more than two detectors with
arbitrary arm directions.Comment: 28 pages, 17 figures, Replaced with the version to be published in
Physical Review
Split luciferase complementation assay to detect regulated protein-protein interactions in rice protoplasts in a large-scale format
BACKGROUND: The rice interactome, in which a network of protein-protein interactions has been elucidated in rice, is a useful resource to identify functional modules of rice signal transduction pathways. Protein-protein interactions occur in cells in two ways, constitutive and regulative. While a yeast-based high-throughput method has been widely used to identify the constitutive interactions, a method to detect the regulated interactions is rarely developed for a large-scale analysis. RESULTS: A split luciferase complementation assay was applied to detect the regulated interactions in rice. A transformation method of rice protoplasts in a 96-well plate was first established for a large-scale analysis. In addition, an antibody that specifically recognizes a carboxyl-terminal fragment of Renilla luciferase was newly developed. A pair of antibodies that recognize amino- and carboxyl- terminal fragments of Renilla luciferase, respectively, was then used to monitor quality and quantity of interacting recombinant-proteins accumulated in the cells. For a proof-of-concept, the method was applied to detect the gibberellin-dependent interaction between GIBBERELLIN INSENSITIVE DWARF1 and SLENDER RICE 1. CONCLUSIONS: A method to detect regulated protein-protein interactions was developed towards establishment of the rice interactome
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