Interplay of the two ancient metabolites auxin and MEcPP regulates adaptive growth.

The ancient morphoregulatory hormone auxin dynamically realigns dedicated cellular processes that shape plant growth under prevailing environmental conditions. However, the nature of the stress-responsive signal altering auxin homeostasis remains elusive. Here we establish that the evolutionarily conserved plastidial retrograde signaling metabolite methylerythritol cyclodiphosphate (MEcPP) controls adaptive growth by dual transcriptional and post-translational regulatory inputs that modulate auxin levels and distribution patterns in response to stress. We demonstrate that in vivo accumulation or exogenous application of MEcPP alters the expression of two auxin reporters, DR5:GFP and DII-VENUS, and reduces the abundance of the auxin-efflux carrier PIN-FORMED1 (PIN1) at the plasma membrane. However, pharmacological intervention with clathrin-mediated endocytosis blocks the PIN1 reduction. This study provides insight into the interplay between these two indispensable signaling metabolites by establishing the mode of MEcPP action in altering auxin homeostasis, and as such, positioning plastidial function as the primary driver of adaptive growth

FAST observations of an extremely active episode of FRB 20201124A: II. Energy Distribution

We report the properties of more than 800 bursts detected from the repeating fast radio burst (FRB) source FRB 20201124A with the Five-hundred-meter Aperture Spherical radio Telescope (FAST) during an extremely active episode on UTC September 25-28, 2021 in a series of four papers. In this second paper of the series, we mainly focus on the energy distribution of the detected bursts. The event rate initially increased exponentially but the source activity stopped within 24 hours after the 4th day. The detection of 542 bursts in one hour during the fourth day marked the highest event rate detected from one single FRB source so far. The bursts have complex structures in the time-frequency space. We find a double-peak distribution of the waiting time, which can be modeled with two log-normal functions peaking at 51.22 ms and 10.05 s, respectively. Compared with the emission from a previous active episode of the source detected with FAST, the second distribution peak time is smaller, suggesting that this peak is defined by the activity level of the source. We calculate the isotropic energy of the bursts using both a partial bandwidth and a full bandwidth and find that the energy distribution is not significantly changed. We find that an exponentially connected broken-power-law function can fit the cumulative burst energy distribution well, with the lower and higher-energy indices being $-1.22\pm0.01$ and $-4.27\pm0.23$, respectively. Assuming a radio radiative efficiency of $\eta_r = 10^{-4}$, the total isotropic energy of the bursts released during the four days when the source was active is already $3.9\times10^{46}$ erg, exceeding $\sim 23\%$ of the available magnetar dipolar magnetic energy. This challenges the magnetar models invoking an inefficient radio emission (e.g. synchrotron maser models).Comment: 26 pages, 7 figures, accepted for publication in Research in Astronomy and Astrophysic

FAST observations of an extremely active episode of FRB 20201124A: IV. Spin Period Search

We report the properties of more than 800 bursts detected from the repeating fast radio burst (FRB) source FRB 20201124A with the Five-hundred-meter Aperture Spherical radio telescope (FAST) during an extremely active episode on UTC September 25th-28th, 2021 in a series of four papers. In this fourth paper of the series, we present a systematic search of the spin period and linear acceleration of the source object from both 996 individual pulse peaks and the dedispersed time series. No credible spin period was found from this data set. We rule out the presence of significant periodicity in the range between 1 ms to 100 s with a pulse duty cycle $< 0.49\pm0.08$ (when the profile is defined by a von-Mises function, not a boxcar function) and linear acceleration up to $300$ m s$^{-2}$ in each of the four one-hour observing sessions, and up to $0.6$ m s$^{-2}$ in all 4 days. These searches contest theoretical scenarios involving a 1 ms to 100 s isolated magnetar/pulsar with surface magnetic field $<10^{15}$ G and a small duty cycle (such as in a polar-cap emission mode) or a pulsar with a companion star or black hole up to 100 M$_{\rm \odot}$ and $P_b>10$ hours. We also perform a periodicity search of the fine structures and identify 53 unrelated millisecond-timescale "periods" in multi-components with the highest significance of 3.9 $\sigma$. The "periods" recovered from the fine structures are neither consistent nor harmonically related. Thus they are not likely to come from a spin period. We caution against claiming spin periodicity with significance below $\sim$ 4 $\sigma$ with multi-components from one-off FRBs. We discuss the implications of our results and the possible connections between FRB multi-components and pulsar micro-structures.Comment: Accepted by Research in Astronomy and Astrophysics (RAA

Crystal structure of rhodopsin bound to arrestin by femtosecond X-ray laser.

G-protein-coupled receptors (GPCRs) signal primarily through G proteins or arrestins. Arrestin binding to GPCRs blocks G protein interaction and redirects signalling to numerous G-protein-independent pathways. Here we report the crystal structure of a constitutively active form of human rhodopsin bound to a pre-activated form of the mouse visual arrestin, determined by serial femtosecond X-ray laser crystallography. Together with extensive biochemical and mutagenesis data, the structure reveals an overall architecture of the rhodopsin-arrestin assembly in which rhodopsin uses distinct structural elements, including transmembrane helix 7 and helix 8, to recruit arrestin. Correspondingly, arrestin adopts the pre-activated conformation, with a ∼20° rotation between the amino and carboxy domains, which opens up a cleft in arrestin to accommodate a short helix formed by the second intracellular loop of rhodopsin. This structure provides a basis for understanding GPCR-mediated arrestin-biased signalling and demonstrates the power of X-ray lasers for advancing the frontiers of structural biology

The Suppressor of AAC2 Lethality SAL1 Modulates Sensitivity of Heterologously Expressed Artemia ADP/ATP Carrier to Bongkrekate in Yeast

The ADP/ATP carrier protein (AAC) expressed in Artemia franciscana is refractory to bongkrekate. We generated two strains of Saccharomyces cerevisiae where AAC1 and AAC3 were inactivated and the AAC2 isoform was replaced with Artemia AAC containing a hemagglutinin tag (ArAAC-HA). In one of the strains the suppressor of ΔAAC2 lethality, SAL1, was also inactivated but a plasmid coding for yeast AAC2 was included, because the ArAACΔsal1Δ strain was lethal. In both strains ArAAC-HA was expressed and correctly localized to the mitochondria. Peptide sequencing of ArAAC expressed in Artemia and that expressed in the modified yeasts revealed identical amino acid sequences. The isolated mitochondria from both modified strains developed 85% of the membrane potential attained by mitochondria of control strains, and addition of ADP yielded bongkrekate-sensitive depolarizations implying acquired sensitivity of ArAAC-mediated adenine nucleotide exchange to this poison, independent from SAL1. However, growth of ArAAC-expressing yeasts in glycerol-containing media was arrested by bongkrekate only in the presence of SAL1. We conclude that the mitochondrial environment of yeasts relying on respiratory growth conferred sensitivity of ArAAC to bongkrekate in a SAL1-dependent manner. © 2013 Wysocka-Kapcinska et al

Metal-organic framework templated electrodeposition of functional gold nanostructures

Utilizing a pair of quick, scalable electrochemical processes, the permanently porous MOF HKUST-1 was electrochemically grown on a copper electrode and this HKUST-1-coated electrode was used to template electrodeposition of a gold nanostructure within the pore network of the MOF. Transmission electron microscopy demonstrates that a proportion of the gold nanostructures exhibit structural features replicating the pore space of this ∼1.4 nm maximum pore diameter MOF, as well as regions that are larger in size. Scanning electron microscopy shows that the electrodeposited gold nanostructure, produced under certain conditions of synthesis and template removal, is sufficiently inter-grown and mechanically robust to retain the octahedral morphology of the HKUST-1 template crystals. The functionality of the gold nanostructure within the crystalline HKUST-1 was demonstrated through the surface enhanced Raman spectroscopic (SERS) detection of 4-fluorothiophenol at concentrations as low as 1 μM. The reported process is confirmed as a viable electrodeposition method for obtaining functional, accessible metal nanostructures encapsulated within MOF crystals

The role of a new CD44st in increasing the invasion capability of the human breast cancer cell line MCF-7

<p>Abstract</p> <p>Background</p> <p>CD44, a hyaluronan (HA) receptor, is a multistructural and multifunctional cell surface molecule involved in cell proliferation, cell differentiation, cell migration, angiogenesis, presentation of cytokines, chemokines and growth factors to the corresponding receptors, and docking of proteases at the cell membrane, as well as in signaling for cell survival. The CD44 gene contains 20 exons that are alternatively spliced, giving rise to many CD44 isoforms, perhaps including tumor-specific sequences.</p> <p>Methods</p> <p>Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to detect CD44st mRNA and CD44 protein in sensitive MCF-7, Lovo, K562 and HL-60 cell lines as well as their parental counterparts, respectively. The full length cDNA encoding CD44st was obtained from the total RNA isolated from MCF-7/Adr cells by RT-PCR, and subcloned into the pMD19-T vector. The CD44st gene sequence and open reading frame were confirmed by restriction enzyme analysis and nucleotide sequencing, and then inserted into the eukaryotic expression vector pcDNA3.1. The pcDNA3.1-CD44st was transfected into MCF-7 cells using Lipofectamine. After transfection, the positive clones were obtained by G418 screening. The changes of the MMP-2 and MMP-9 genes and protein levels were detected by RT-PCR and gelatin zymography, respectively. The number of the cells penetrating through the artificial matrix membrane in each group (MCF-7, MCF-7+HA, MCF-7/neo, MCF-7/neo+HA, MCF-7/CD44st, MCF-7/CD44st+HA and MCF-7/CD44st+Anti-CD44+HA) was counted to compare the change of the invasion capability regulated by the CD44st. Erk and P-Erk were investigated by Western blotting to approach the molecular mechanisms of MMP-2 and MMP-9 expression regulated by the CD44st.</p> <p>Results</p> <p>Sensitive MCF-7, Lovo, K562 and HL-60 cells did not contain CD44st mRNA and CD44 protein. In contrast, the multidrug resistance MCF-7/Adr, Lovo/Adr, K562/Adr and HL-60/Adr cells expressed CD44st mRNA and CD44 protein. The CD44st mRNA gene sequence was successfully cloned into the recombinant vector pcDNA3.1 and identified by the two restriction enzymes. It was confirmed that the reconstructed plasmid contained the gene sequence of CD44st that was composed of exons 1 to 4, 16 to 17, and 1 to 205 bases of exons 18. The new gene sequence was sent to NCBI for publication, and obtained the registration number FJ216964. The up-regulated level of the mRNA of the CD44 gene and the CD44 protein were detected, respectively, by RT-PCR and flow cytometry in MCF-7 cells transfected with pcDNA3.1-CD44st. The invasiveness of the cells and the activity of MMP-2 and MMP-9 were clearly activated by HA treatment, and blocked by CD44 neutralizing antibody. MCF-7/CD44st cells pretreated with the neutralizing antibody against CD44, and the inhibitor of MAPKs signaling pathway, could strongly block the expression of P-Erk.</p> <p>Conclusions</p> <p>A new CD44st was expressed in multidrug resistant MCF-7/Adr, Lovo/Adr, K562/Adr and HL-60/Adr cells. The expression vector pcDNA3.1-CD44st was cloned and constructed successfully, and stably transfected into MCF-7 cells. HA could interact with the new CD44st and regulate the expression of MMP-2 and MMP-9, which could increase the invasion capability of MCF-7 cells through the Ras/MAPK signaling pathway.</p

FAST observations of an extremely active episode of FRB 20201124A: III. Polarimetry

As the third paper in the multiple-part series, we report the statistical properties of radio bursts detected from the repeating fast radio burst (FRB) source FRB 20201124A with the Five-hundred-meter Aperture Spherical radio telescope (FAST) during an extremely active episode between the 25th and the 28th of September 2021 (UT). We focus on the polarisation properties of 536 bright bursts with $\mathrm{S/N}>50$. We found that the Faraday rotation measures (RMs) monotonically dropped from $-579 \ {\rm rad \ m^{-2}}$ to $-605 \ {\rm rad \ m^{-2}}$ in the 4-day window. The RM values were compatible with the values ($-300$ to $-900\ {\rm rad \ m^{-2}}$ ) reported 4 month ago (Xu et al. 2022). However, the RM evolution rate in the current observation window was at least an order of magnitude smaller than the one ($\sim 500\ {\rm rad \ m^{-2}\, day^{-1}}$) previously reported during the rapid RM-variation phase, but is still higher than the one ($\le 1\ {\rm rad \ m^{-2} day^{-1}}$) during the later RM no-evolution phase. The bursts of FRB 20201124A were highly polarised with the total degree of polarisation (circular plus linear) greater than 90polarisation position angles (PAs), degree of linear polarisation ($L/I$), and degree of circular polarisation ($V/I$) can be characterised with unimodal distribution functions. During the observation window, the distributions became wider with time, i.e. with larger scatter, but the centroids of the distribution functions remained nearly constant. For individual bursts, significant PA variations (confidence level 5-$\sigma$) were observed in 33% of all bursts. The polarisation of single pulses seems to follow certain complex trajectories on the Poincar\'e sphere, which may shed light on the radiation mechanism at the source or the plasma properties along the path of FRB propagation.Comment: 25 pages, 16 figures. Accepted by Research in Astronomy and Astrophysics (RAA