1,349 research outputs found
Two-step membrane binding by the bacterial SRP receptor enable efficient and accurate Co-translational protein targeting
The signal recognition particle (SRP) delivers ~30% of the proteome to the eukaryotic endoplasmic reticulum, or the bacterial plasma membrane. The precise mechanism by which the bacterial SRP receptor, FtsY, interacts with and is regulated at the target membrane remain unclear. Here, quantitative analysis of FtsY-lipid interactions at single-molecule resolution revealed a two-step mechanism in which FtsY initially contacts membrane via a Dynamic mode, followed by an SRP-induced conformational transition to a Stable mode that activates FtsY for downstream steps. Importantly, mutational analyses revealed extensive auto-inhibitory mechanisms that prevent free FtsY from engaging membrane in the Stable mode; an engineered FtsY pre-organized into the Stable mode led to indiscriminate targeting in vitro and disrupted FtsY function in vivo. Our results show that the two-step lipid-binding mechanism uncouples the membrane association of FtsY from its conformational activation, thus optimizing the balance between the efficiency and fidelity of co-translational protein targeting
Metrology Camera System of Prime Focus Spectrograph for Subaru Telescope
The Prime Focus Spectrograph (PFS) is a new optical/near-infrared multi-fiber
spectrograph designed for the prime focus of the 8.2m Subaru telescope. PFS
will cover a 1.3 degree diameter field with 2394 fibers to complement the
imaging capabilities of Hyper SuprimeCam. To retain high throughput, the final
positioning accuracy between the fibers and observing targets of PFS is
required to be less than 10um. The metrology camera system (MCS) serves as the
optical encoder of the fiber motors for the configuring of fibers. MCS provides
the fiber positions within a 5um error over the 45 cm focal plane. The
information from MCS will be fed into the fiber positioner control system for
the closed loop control. MCS will be located at the Cassegrain focus of Subaru
telescope in order to to cover the whole focal plane with one 50M pixel Canon
CMOS camera. It is a 380mm Schmidt type telescope which generates a uniform
spot size with a 10 micron FWHM across the field for reasonable sampling of
PSF. Carbon fiber tubes are used to provide a stable structure over the
operating conditions without focus adjustments. The CMOS sensor can be read in
0.8s to reduce the overhead for the fiber configuration. The positions of all
fibers can be obtained within 0.5s after the readout of the frame. This enables
the overall fiber configuration to be less than 2 minutes. MCS will be
installed inside a standard Subaru Cassgrain Box. All components that generate
heat are located inside a glycol cooled cabinet to reduce the possible image
motion due to heat. The optics and camera for MCS have been delivered and
tested. The mechanical parts and supporting structure are ready as of spring
2016. The integration of MCS will start in the summer of 2016.Comment: 11 pages, 15 figures. SPIE proceeding. arXiv admin note: text overlap
with arXiv:1408.287
Fluctuating magnetic droplets immersed in a sea of quantum spin liquid
The search of quantum spin liquid (QSL), an exotic magnetic state with
strongly-fluctuating and highly-entangled spins down to zero temperature, is a
main theme in current condensed matter physics. However, there is no
smoking-gun evidence for deconfined spinons in any QSL candidate so far. The
disorders and competing exchange interactions may prevent the formation of an
ideal QSL state on frustrated spin lattices. Here we report comprehensive and
systematic measurements of the magnetic susceptibility, ultra-low temperature
specific heat, muon spin relaxation (muSR), nuclear magnetic resonance (NMR),
and thermal conductivity for NaYbSe2 single crystals, in which Yb3+ ions with
effective spin-1/2 form a perfect triangular lattice. All these complementary
techniques find no evidence of long-range magnetic order down to their
respective base temperatures. Instead, specific heat, muSR and NMR measurements
suggest the coexistence of quasi-static and dynamic spins in NaYbSe2. The
scattering from these quasi-static spins may cause the absence of magnetic
thermal conductivity. Thus, we propose a scenario of fluctuating ferrimagnetic
droplets immersed in a sea of QSL. This may be quite common on the way pursuing
an ideal QSL, and provides a brand-new platform to study how a QSL state
survives impurities and coexists with other magnetically ordered states
Identification of novel DNA methylation inhibitors via a two-component reporter gene system
<p>Abstract</p> <p>Background</p> <p>Targeting abnormal DNA methylation represents a therapeutically relevant strategy for cancer treatment as demonstrated by the US Food and Drug Administration approval of the DNA methyltransferase inhibitors azacytidine and 5-aza-2'-deoxycytidine for the treatment of myelodysplastic syndromes. But their use is associated with increased incidences of bone marrow suppression. Alternatively, procainamide has emerged as a potential DNA demethylating agent for clinical translation. While procainamide is much safer than 5-aza-2'-deoxycytidine, it requires high concentrations to be effective in DNA demethylation in suppressing cancer cell growth. Thus, our laboratories have embarked on the pharmacological exploitation of procainamide to develop potent DNA methylation inhibitors through lead optimization.</p> <p>Methods</p> <p>We report the use of a DNA methylation two-component enhanced green fluorescent protein reporter system as a screening platform to identify novel DNA methylation inhibitors from a compound library containing procainamide derivatives.</p> <p>Results</p> <p>A lead agent IM25, which exhibits substantially higher potency in <it>GSTp1 </it>DNA demethylation with lower cytotoxicity in MCF7 cells relative to procainamide and 5-aza-2'-deoxycytidine, was identified by the screening platform.</p> <p>Conclusions</p> <p>Our data provide a proof-of-concept that procainamide could be pharmacologically exploited to develop novel DNA methylation inhibitors, of which the translational potential in cancer therapy/prevention is currently under investigation.</p
Hypermethylation of the TGF-β target, ABCA1 is associated with poor prognosis in ovarian cancer patients
Background
The dysregulation of transforming growth factor-β (TGF-β) signaling plays a crucial role in ovarian carcinogenesis and in maintaining cancer stem cell properties. Classified as a member of the ATP-binding cassette (ABC) family, ABCA1 was previously identified by methylated DNA immunoprecipitation microarray (mDIP-Chip) to be methylated in ovarian cancer cell lines, A2780 and CP70. By microarray, it was also found to be upregulated in immortalized ovarian surface epithelial (IOSE) cells following TGF-β treatment. Thus, we hypothesized that ABCA1 may be involved in ovarian cancer and its initiation.
Results
We first compared the expression level of ABCA1 in IOSE cells and a panel of ovarian cancer cell lines and found that ABCA1 was expressed in HeyC2, SKOV3, MCP3, and MCP2 ovarian cancer cell lines but downregulated in A2780 and CP70 ovarian cancer cell lines. The reduced expression of ABCA1 in A2780 and CP70 cells was associated with promoter hypermethylation, as demonstrated by bisulfite pyro-sequencing. We also found that knockdown of ABCA1 increased the cholesterol level and promoted cell growth in vitro and in vivo. Further analysis of ABCA1 methylation in 76 ovarian cancer patient samples demonstrated that patients with higher ABCA1 methylation are associated with high stage (P = 0.0131) and grade (P = 0.0137). Kaplan-Meier analysis also found that patients with higher levels of methylation of ABCA1 have shorter overall survival (P = 0.019). Furthermore, tissue microarray using 55 ovarian cancer patient samples revealed that patients with a lower level of ABCA1 expression are associated with shorter progress-free survival (P = 0.038).
Conclusions
ABCA1 may be a tumor suppressor and is hypermethylated in a subset of ovarian cancer patients. Hypermethylation of ABCA1 is associated with poor prognosis in these patients
Dirac semimetal PdTe2 temperature-dependent quasiparticle dynamics and electron-phonon coupling
Dirac semimetal PdTe2 single-crystal temperature-dependent ultrafast carrier
and phonon dynamics were studied using ultrafast optical pump-probe
spectroscopy. Two distinct carrier and coherent phonons relaxation processes
were identified in the 5 K - 300 K range. Quantitative analysis revealed a fast
relaxation process ({\tau}_f) occurring on a subpicosecond time scale which
originated from electron-phonon thermalization. This was followed by a slower
relaxation process ({\tau}_s) with a time scale of ~ 7-9.5 ps which originated
from phonon-assisted electron-hole recombination. Two significant vibrational
modes resolved at all measured temperatures and corresponded to Te atoms
in-plane (E_g), and out-of-plane (A_1g), motion. As temperature increased both
phonon modes softened markedly. A_1g mode frequency monotonically decreased as
temperature increased. Its damping rate remained virtually unchanged. As
expected, E_g decreased uniformly as temperatures rose. At temperatures above
80 K, there was insignificant change. Test results suggested that pure
dephasing played an important role in the relaxation processes. PdTe2 phonon is
thought responsible for its superconductive properties. Examining phonons
behavior should improve the understanding of its complex superconductivity.Comment: 6 pages, 4 figure
The Epigenetic Modifier PRDM5 Functions as a Tumor Suppressor through Modulating WNT/β-Catenin Signaling and Is Frequently Silenced in Multiple Tumors
BACKGROUND: PRDM (PRDI-BF1 and RIZ domain containing) proteins are zinc finger proteins involved in multiple cellular regulations by acting as epigenetic modifiers. We studied a recently identified PRDM member PRDM5 for its epigenetic abnormality and tumor suppressive functions in multiple tumorigeneses. METHODOLOGY/PRINCIPAL FINDINGS: Semi-quantitative RT-PCR showed that PRDM5 was broadly expressed in human normal tissues, but frequently silenced or downregulated in multiple carcinoma cell lines due to promoter CpG methylation, including 80% (4/5) nasopharyngeal, 44% (8/18) esophageal, 76% (13/17) gastric, 50% (2/4) cervical, and 25% (3/12) hepatocellular carcinoma cell lines, but not in any immortalized normal epithelial cell lines. PRDM5 expression could be restored by 5-aza-2'-deoxycytidine demethylation treatment in silenced cell lines. PRDM5 methylation was frequently detected by methylation-specific PCR (MSP) in multiple primary tumors, including 93% (43/46) nasopharyngeal, 58% (25/43) esophageal, 88% (37/42) gastric and 63% (29/46) hepatocellular tumors. PRDM5 was further found a stress-responsive gene, but its response was impaired when the promoter was methylated. Ectopic PRDM5 expression significantly inhibited tumor cell clonogenicity, accompanied by the inhibition of TCF/β-catenin-dependent transcription and downregulation of CDK4, TWIST1 and MDM2 oncogenes, while knocking down of PRDM5 expression lead to increased cell proliferation. ChIP assay showed that PRDM5 bound to its target gene promoters and suppressed their transcription. An inverse correlation between the expression of PRDM5 and activated β-catenin was also observed in cell lines. CONCLUSIONS/SIGNIFICANCE: PRDM5 functions as a tumor suppressor at least partially through antagonizing aberrant WNT/β-catenin signaling and oncogene expression. Frequent epigenetic silencing of PRDM5 is involved in multiple tumorigeneses, which could serve as a tumor biomarker
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