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The PIWI-interacting RNA (piRNA) pathway plays a crucial role in silencing transposons in the germline. piRNA-guided target cleavage by PIWI proteins triggers the biogenesis of new piRNAs from the cleaved RNA fragments. This process, known as the ping-pong cycle, is mediated by the two PIWI proteins, Siwi and BmAgo3, in silkworms. However, the detailed molecular mechanism of the ping-pong cycle remains largely unclear. Here, we show that Spindle-E (Spn-E), a putative ATP-dependent RNA helicase, is essential for BmAgo3-dependent production of Siwi-bound piRNAs in the ping-pong cycle and that this function of Spn-E requires its ATPase activity. Moreover, Spn-E acts to suppress homotypic Siwi–Siwi ping-pong, but this function of Spn-E is independent of its ATPase activity. These results highlight the dual role of Spn-E in facilitating proper heterotypic ping-pong in silkworms. Synopsis: Spn-E promotes BmAgo3-dependent production of Siwi-bound piRNAs using its ATPase activity while suppressing Siwi-Siwi homotypic ping-pong, thereby facilitating proper heterotypic ping-pong in silkworms. Spn-E promotes proper heterotypic ping-pong between Siwi and BmAgo3 in silkworms. Spn-E's ATPase activity is required for BmAgo3-dependent production of Siwi-bound piRNAs. Spn-E suppresses Siwi-Siwi homotypic ping-pong independently of its ATPase activity. Spn-E promotes BmAgo3-dependent production of Siwi-bound piRNAs using its ATPase activity while suppressing Siwi-Siwi homotypic ping-pong, thereby facilitating proper heterotypic ping-pong in silkworms. [ABSTRACT FROM AUTHOR]

Fragmentation during the early stages of high-mass star formation is crucial for understanding the formation of high-mass clusters. We investigated fragmentation within 39 high-mass star-forming clumps as part of the Atacama Large Millimeter/submillimeter Array Survey of 70 μ m Dark High-mass Clumps in Early Stages (ASHES) survey. Considering projection effects, we have estimated core separations for 839 cores identified from the continuum emission and found mean values between 0.08 and 0.32 pc within each clump. We find compatibility of the observed core separations and masses with the thermal Jeans length and mass, respectively. We also present subclump structures revealed by the 7 m array continuum emission. Comparison of the Jeans parameters using clump and subclump densities with the separation and masses of gravitationally bound cores suggests that they can be explained by clump fragmentation, implying the simultaneous formation of subclumps and cores within rather than a step-by-step hierarchical fragmentation. The number of cores in each clump positively correlates with the clump surface density and the number expected from the thermal Jeans fragmentation. We also find that the higher the fraction of protostellar cores, the larger the dynamic range of the core mass, implying that the cores are growing in mass as the clump evolves. The ASHES sample exhibits various fragmentation patterns: aligned, scattered, clustered, and subclustered. Using the Q -parameter, which can help distinguish between centrally condensed and subclustered spatial core distributions, we finally find that in the early evolutionary stages of high-mass star formation, cores tend to follow a subclustered distribution. [ABSTRACT FROM AUTHOR]

We examine spatial variations of the C0 |$/$| CO abundance ratio (X C/CO) in the vicinity of the γ-ray supernova remnant W 51 C, based on [C  i ] (3 P 1–3 P 0), 12CO(1–0), and 13CO(1–0) observations with the ASTE and Nobeyama 45 m telescopes. We find that X C/CO varies in a range of 0.02–0.16 (0.05 in median) inside the molecular clouds of A V > 100 mag, where photodissociation of CO by the interstellar UV is negligible. Furthermore, X C/CO is locally enhanced by a factor of up to four near the W 51 C center, depending on the projected distance from the W 51 C center. In high- A V molecular clouds, X C/CO is determined by the ratio of the cosmic-ray (CR) ionization rate to the H2 density, and we find no clear spatial variation of the H2 density against the projected distance. Hence, the high CR ionization rate may locally enhance X C/CO near the W 51 C center. We also find that the observed spatial extent of the enhanced X C/CO (∼17 pc) is consistent with the diffusion distance of CRs with an energy of 100 MeV. This fact suggests that the low-energy CRs accelerated in W 51 C enhance X C/CO. The CR ionization rate in the X C/CO-enhanced cloud is estimated to be 3 × 10−16 s−1 on the basis of time-dependent photodissociation region simulations of X C/CO, the value of which is 30 times higher than that in the standard Galactic environment. These results demonstrate that [C  i ] is a powerful probe to investigate the interaction between CRs and the interstellar medium for a wide area in the vicinity of supernova remnants. [ABSTRACT FROM AUTHOR]

PIWI-interacting RNAs (piRNAs) guide PIWI proteins to target transposons in germline cells, thereby suppressing transposon activity to preserve genome integrity in metazoans' gonadal tissues. Piwi, one of three Drosophila PIWI proteins, is expressed in the nucleus and suppresses transposon activity by forming heterochromatin in an RNA cleavage-independent manner. Recently, Piwi was reported to control cell metabolism in Drosophila fat body, providing an example of piRNAs acting in non-gonadal somatic tissues. However, mutant flies of the other two PIWI proteins, Aubergine (Aub) and Argonaute3 (Ago3), show no apparent phenotype except for infertility, blurring the importance of the piRNA pathway in non-gonadal somatic tissues. The silkworm, Bombyx mori, possesses two PIWI proteins, Siwi (Aub homolog) and BmAgo3 (Ago3 homolog), whereas B. mori does not have a Piwi homolog. Siwi and BmAgo3 are mainly expressed in gonadal tissues and play a role in repressing transposon activity by cleaving transposon RNA in the cytoplasm. Here, we generated Siwi and BmAgo3 loss-of-function mutants of B. mori and found that they both showed delayed larval growth and failed to become adult moths. They also exhibited defects in wing development and sexual differentiation. Transcriptome analysis revealed that loss of somatic piRNA biogenesis pathways results in abnormal expression of not only transposons but also host genes, presumably causing severe growth defects. Our results highlight the roles of non-gonadal somatic piRNAs in B. mori development. Author summary: In animal germ cells, PIWI proteins and piRNAs play an important role in the defense system against selfish elements such as transposons. The fruit fly Drosophila possesses two PIWI proteins, Aub and Ago3, both of which are required for piRNA production and transposon repression in the cytoplasm of germ cells. However, their roles in somatic cells are unclear. Using a lepidopteran model insect, the silkworm Bombyx mori, we established Siwi (Aub homolog) and BmAgo3 (Ago3 homolog) germline knockout mutants and characterized non-gonadal somatic piRNA functions in B. mori development. We found that somatic piRNAs are utilized to suppress transposon activity and maintain normal gene expression in several somatic tissues. Unlike Drosophila, disruption of somatic piRNA pathways severely inhibited normal development in somatic tissues of B. mori: both mutants grew slowly, did not develop to adult moths, and exhibited defects in wing development and sexual differentiation. This study is the first to generate germline knockout insects of PIWI genes in non-Drosophila insects and expands the knowledge on the ancestral or universal functions of piRNAs in arthropods. [ABSTRACT FROM AUTHOR]

Studying the physical and chemical properties of cold and dense molecular clouds is crucial for the understanding of how stars form. Under the typical conditions of infrared dark clouds, CO is removed from the gas phase and trapped onto the surface of dust grains by the so-called depletion process. This suggests that the CO-depletion factor (f D) can be a useful chemical indicator for identifying cold and dense regions (i.e., prestellar cores). We have used the 1.3 mm continuum and C18O (2â€"1) data observed at the resolution of ∼5000 au in the ALMA Survey of 70 ÎĽ m Dark High-mass Clumps in Early Stages (ASHES) to construct averaged maps of f D in 12 clumps to characterize the earliest stages of the high-mass star formation process. The average f D determined for 277 of the 294 ASHES cores follows an unexpected increase from the prestellar to the protostellar stage. If we exclude the temperature effect due to the slight variations in the NH3 kinetic temperature among different cores, we explain this result as a dependence primarily on the average gas density, which increases in cores where protostellar conditions prevail. This shows that f D determined in high-mass star-forming regions at the core scale is insufficient to distinguish among prestellar and protostellar conditions for the individual cores and should be complemented by information provided by additional tracers. However, we confirm that the clump-averaged f D values correlate with the luminosity-to-mass ratio of each source, which is known to trace the evolution of the star formation process. [ABSTRACT FROM AUTHOR]

Argonaute proteins use nucleic acid guides to find and bind specific DNA or RNA target sequences. Argonaute proteins have diverse biological functions and many retain their ancestral endoribonuclease activity, cleaving the phosphodiester bond between target nucleotides t10 and t11. In animals, the PIWI proteins—a specialized class of Argonaute proteins—use 21–35 nucleotide PIWI-interacting RNAs (piRNAs) to direct transposon silencing, protect the germline genome, and regulate gene expression during gametogenesis1. The piRNA pathway is required for fertility in one or both sexes of nearly all animals. Both piRNA production and function require RNA cleavage catalysed by PIWI proteins. Spermatogenesis in mice and other placental mammals requires three distinct, developmentally regulated PIWI proteins: MIWI (PIWIL1), MILI (PIWIL2) and MIWI22–4 (PIWIL4). The piRNA-guided endoribonuclease activities of MIWI and MILI are essential for the production of functional sperm5,6. piRNA-directed silencing in mice and insects also requires GTSF1, a PIWI-associated protein of unknown function7–12. Here we report that GTSF1 potentiates the weak, intrinsic, piRNA-directed RNA cleavage activities of PIWI proteins, transforming them into efficient endoribonucleases. GTSF1 is thus an example of an auxiliary protein that potentiates the catalytic activity of an Argonaute protein.The evolutionarily conserved RNA-binding protein GTSF1 and its homologues interact with members of the PIWI class of Argonaute proteins, increasing the efficiency of the RNA-cleaving activity of PIWI proteins, an essential function across the animal kingdom. [ABSTRACT FROM AUTHOR]

With a mass of ∼1000 M ⊙ and a surface density of ∼0.5 g cmâˆ'2, G023.477+0.114, also known as IRDC 18310-4, is an infrared dark cloud (IRDC) that has the potential to form high-mass stars and has been recognized as a promising prestellar clump candidate. To characterize the early stages of high-mass star formation, we have observed G023.477+0.114 as part of the Atacama Large Millimeter/submillimeter Array (ALMA) Survey of 70 ÎĽ m Dark High-mass Clumps in Early Stages. We have conducted ∼1.″2 resolution observations with ALMA at 1.3 mm in dust continuum and molecular line emission. We have identified 11 cores, whose masses range from 1.1 to 19.0 M ⊙. Ignoring magnetic fields, the virial parameters of the cores are below unity, implying that the cores are gravitationally bound. However, when magnetic fields are included, the prestellar cores are close to virial equilibrium, while the protostellar cores remain sub-virialized. Star formation activity has already started in this clump. Four collimated outflows are detected in CO and SiO. H2CO and CH3OH emission coincide with the high-velocity components seen in the CO and SiO emission. The outflows are randomly oriented for the natal filament and the magnetic field. The position-velocity diagrams suggest that episodic mass ejection has already begun even in this very early phase of protostellar formation. The masses of the identified cores are comparable to the expected maximum stellar mass that this IRDC could form (8â€"19 M ⊙). We explore two possibilities on how IRDC G023.477+0.114 could eventually form high-mass stars in the context of theoretical scenarios. [ABSTRACT FROM AUTHOR]

In animal germlines, PIWI proteins and the associated PIWI-interacting RNAs (piRNAs) protect genome integrity by silencing transposons. Here we report the extensive sequence and quantitative correlations between 2′,3′-cyclic phosphate-containing RNAs (cP-RNAs), identified using cP-RNA-seq, and piRNAs in the Bombyx germ cell line and mouse testes. The cP-RNAs containing 5′-phosphate (P-cP-RNAs) identified by P-cP-RNA-seq harbor highly consistent 5′-end positions as the piRNAs and are loaded onto PIWI protein, suggesting their direct utilization as piRNA precursors. We identified Bombyx RNase Kappa (BmRNase κ) as a mitochondria-associated endoribonuclease which produces cP-RNAs during piRNA biogenesis. BmRNase κ-depletion elevated transposon levels and disrupted a piRNA-mediated sex determination in Bombyx embryos, indicating the crucial roles of BmRNase κ in piRNA biogenesis and embryonic development. Our results reveal a BmRNase κ-engaged piRNA biogenesis pathway, in which the generation of cP-RNAs promotes robust piRNA production. Mature piRNAs are produced from long single-stranded precursor RNAs. Here the authors show that RNase κ generates 2′,3′-cyclic phosphate-containing piRNA precursors and promotes piRNA production. [ABSTRACT FROM AUTHOR]

We present a spatio-kinematical analysis of the CO (J = 2 → 1) line emission, observed with the Atacama Large Millimeter/submillimeter Array (ALMA), of the outflow associated with the most massive core, ALMA1, in the 70 μm dark clump G010.991–00.082. The position–velocity (PV) diagram of the molecular outflow exhibits a peculiar S-shaped morphology that has not been seen in any other star-forming region. We propose a spatio-kinematical model for the bipolar molecular outflow that consists of a decelerating high-velocity component surrounded by a slower component whose velocity increases with distance from the central source. The physical interpretation of the model is in terms of a jet that decelerates as it entrains material from the ambient medium, which has been predicted by calculations and numerical simulations of molecular outflows in the past. One side of the outflow is shorter and shows a stronger deceleration, suggesting that the medium through which the jet moves is significantly inhomogeneous. The age of the outflow is estimated to be τ ≈ 1300 yr, after correction for a mean inclination of the system of ≈57°. [ABSTRACT FROM AUTHOR]

We present observations of the 3 P 1–3 P 0 fine-structure line of atomic carbon using the ASTE 10m sub-mm telescope towards RCW 38, the youngest super star cluster in the Milky Way. The detected [C  i ] emission is compared with the CO J = 1–0 image cube presented in Fukui et al. (2016 , ApJ, 820, 26) which has an angular resolution of 40″ (∼0.33 pc). The overall distribution of the [C  i ] emission in this cluster is similar to that of the 13CO emission. The optical depth of the [C  i ] emission was found to be τ = 0.1–0.6, suggesting mostly optically thin emission. An empirical conversion factor from the [C  i ] integrated intensity to the H2 column density was estimated as X [C  i ] |$= 6.3 \times 10 ^{20}\:$| cm−2 K−1 km−1 s (for visual extinction: A V ≤ 10 mag) and 1.4 × 1021 cm−2 K−1 km−1 s (for A V of 10–100 mag). The column density ratio of the [C  i ] to CO (N [C  i ] |$/N_{\rm CO}$|⁠) was derived as ∼0.1 for A V of 10–100 mag, which is consistent with that of the Orion cloud presented in Ikeda et al. (2002 , ApJ, 571, 560). However, our results cover an A V regime of up to 100 mag, which is wider than the coverage found in Orion, which reaches up to ∼60 mag. Such a high [C  i ] |$/$| CO ratio in a high- A V region is difficult to explain via the plane-parallel photodissociation region model, which predicts that this ratio is close to 0 due to the heavy shielding of the ultraviolet (UV) radiation. Our results suggest that the molecular gas in this cluster is highly clumpy, allowing deep penetration of UV radiation even at averaged A V values of 100 mag. Recent theoretical works have presented models consistent with such clumped gas distribution with a sub-pc clump size (e.g. Tachihara et al. 2018 , arXiv:1811.02224). [ABSTRACT FROM AUTHOR]

We present a study of outflows at extremely early stages of high-mass star formation obtained from the ALMA Survey of 70 μm dark High-mass clumps in Early Stages (ASHES). Twelve massive 3.6−70 μm dark prestellar clump candidates were observed with the Atacama Large Millimeter/submillimeter Array (ALMA) in Band 6. Forty-three outflows are identified toward 41 out of 301 dense cores using the CO and SiO emission lines, yielding a detection rate of 14%. We discover six episodic molecular outflows associated with low- to high-mass cores, indicating that episodic outflows (and therefore episodic accretion) begin at extremely early stages of protostellar evolution for a range of core masses. The time span between consecutive ejection events is much smaller than those found in more evolved stages, which indicates that the ejection episodicity timescale is likely not constant over time. The estimated outflow dynamical timescale appears to increase with core masses, which likely indicates that more massive cores have longer accretion timescales than less massive cores. The lower accretion rates in these 70 μm dark objects compared to the more evolved protostars indicate that the accretion rates increase with time. The total outflow energy rate is smaller than the turbulent energy dissipation rate, which suggests that outflow-induced turbulence cannot sustain the internal clump turbulence at the current epoch. We often detect thermal SiO emission within these 70 μm dark clumps that is unrelated to CO outflows. This SiO emission could be produced by collisions, intersection flows, undetected protostars, or other motions. [ABSTRACT FROM AUTHOR]

We report on measurements of parallax and proper motion for four 22 GHz water maser sources as part of the VERA Outer Rotation Curve project. All the sources show Galactic latitudes of >2° and Galactocentric distances of >11 kpc at the Galactic longitude range of 95° < l < 126°. The sources trace the Galactic warp reaching to 200–400 pc, and also the signature of the warp to 600 pc toward the north Galactic pole. The new results, along with previous results in the literature, show that the maximum height of the Galactic warp increases with Galactocentric distance. Also, we examined velocities perpendicular to the disk for the sample, and found oscillatory behavior between the vertical velocities and Galactic heights. This behavior suggests the existence of bending (vertical density) waves, possibly induced by a perturbing satellite (e.g. the passage of the Sagittarius dwarf galaxy). [ABSTRACT FROM AUTHOR]

We analyze molecular-gas formation in neutral atomic hydrogen (H  i) clouds using the latest CO data, obtained from the FOREST (four-beam receiver system on the 45 m telescope) unbiased Galactic plane imaging survey with the Nobeyama 45 m telescope, and using H  i data taken from the Very Large Array Galactic plane survey. We applied a dendrogram algorithm to the H  i data cube to identify H  i clouds, and we calculated the H  i mass and molecular-gas mass by summing the CO line intensity within each H  i cloud. On the basis of the results, we created a catalog of 5737 identified H  i clouds with local standard of rest (LSR) velocity of V LSR ≤ −20 km s−1 in galactic longitude and latitude ranges of 20° ≤ l ≤ 50° and −1° ≤ b ≤ 1°, respectively. We found that most of the H  i clouds are distributed within a Galactocentric distance of 16 kpc, and most of them are in the cold neutral medium phase. In addition, we determined that the high-mass end of the H  i mass function is fitted well with a power-law function with an index of 2.3. Although two sequences of self-gravitating and diffuse clouds are expected to appear in the M  tot– |$M_{\,{\rm H}_2}$| diagram according to previous works based on a plane-parallel model, the observational data show only a single sequence with large scattering within these two sequences. This implies that most of the clouds are mixtures of these two types of clouds. Moreover, we suggest the following scenario of molecular-gas formation: an H  i -dominant cloud evolved with increasing H2 mass along a path of |$M_{\,{\rm H}_2} \propto M_{\,\rm tot}^2$| by collecting diffuse gas before reaching and moving along the curves of the two sequences. [ABSTRACT FROM AUTHOR]

PIWI-interacting RNAs (piRNAs) of between approximately 24 and 31 nucleotides in length guide PIWI proteins to silence transposons in animal gonads, thereby ensuring fertility1. In the biogenesis of piRNAs, PIWI proteins are first loaded with 5′-monophosphorylated RNA fragments called pre-pre-piRNAs, which then undergo endonucleolytic cleavage to produce pre-piRNAs1,2. Subsequently, the 3′-ends of pre-piRNAs are trimmed by the exonuclease Trimmer (PNLDC1 in mouse)3–6 and 2′-O-methylated by the methyltransferase Hen1 (HENMT1 in mouse)7–9, generating mature piRNAs. It is assumed that the endonuclease Zucchini (MitoPLD in mouse) is a major enzyme catalysing the cleavage of pre-pre-piRNAs into pre-piRNAs10–13. However, direct evidence for this model is lacking, and how pre-piRNAs are generated remains unclear. Here, to analyse pre-piRNA production, we established a Trimmer-knockout silkworm cell line and derived a cell-free system that faithfully recapitulates Zucchini-mediated cleavage of PIWI-loaded pre-pre-piRNAs. We found that pre-piRNAs are generated by parallel Zucchini-dependent and -independent mechanisms. Cleavage by Zucchini occurs at previously unrecognized consensus motifs on pre-pre-piRNAs, requires the RNA helicase Armitage, and is accompanied by 2′-O-methylation of pre-piRNAs. By contrast, slicing of pre-pre-piRNAs with weak Zucchini motifs is achieved by downstream complementary piRNAs, producing pre-piRNAs without 2′-O-methylation. Regardless of the endonucleolytic mechanism, pre-piRNAs are matured by Trimmer and Hen1. Our findings highlight multiplexed processing of piRNA precursors that supports robust and flexible piRNA biogenesis. A silkworm model recapitulates key steps of Zucchini-mediated cleavage of pre-pre-piRNA and provides insights into Zucchini-mediated and -independent pathways that generate pre-piRNAs, which converge to a common piRNA maturation step. [ABSTRACT FROM AUTHOR]

We developed and deployed a simple add-on multi-temperature calibrator for our multicolor transition edge sensor (TES) bolometer camera aimed at simultaneous observation with observing wavelengths of 1.1 and 0.85 mm. To cover the power loading level from the atmospheric emission corresponding to precipitable water vapor (PWV) of 0.5-4 mm, the calibrator consists of spherical mirrors to show the low-temperature stages of the cryostat and filters with moderate opacity to mimic the eight-temperature cold blackbodies. The loading powers introduced by each filter were self-calibrated by measuring the load curves of the TES bolometers when a filter was placed in front of the cryostat window. Each science observation was preceded by the calibration process, which measures the response of the TES bolometers to the atmosphere and filters of various opacities. Then, the responsivities of TES bolometers were derived to convert their output signal to the loading power and correct for the nonlinearity inherent in its response. Furthermore, the loading power falling on the TES bolometers from atmospheric emission measured at various PWV was in good correlation with the PWV measured with the radiometer, which enables the atmospheric extinction correction by fast and sensitive bolometers compared to the available radiometers with the modest sampling speeds. [ABSTRACT FROM AUTHOR]

We developed a simple add-on, cryogen-free, and low-power consumption calibrator for a new transition-edge sensor (TES) bolometer camera mounted on the ASTE 10-m telescope. To measure the responsivity of the TES bolometers and accurately correct for the nonlinearity and atmospheric extinction, we designed a motor-driven rotating filter wheel system installed in front of the cryostat window. This calibrator is required to cover the loading power under various atmospheric conditions, which corresponds to precipitable water vapor (PWV) of 0.5-4 mm. For this range of PWV, 25-100 K blackbodies are necessary for the observing bands of 1.1 and 0.85 mm. To simulate the temperature range, bolometers in the cryostat are also optically coupled to the low-temperature stage (<4 K) inside the cryostat by spherical mirrors. In addition, we used moderately absorptive polystyrene plates that are placed between a spherical mirror and the cryostat window. Various combinations of filters result in eight different temperatures by the filter wheel system and simulate the atmospheric emission under various weather conditions at the ASTE site. [ABSTRACT FROM AUTHOR]

We present a high-resolution (R  ≃ 20  000) near-infrared (9100–13 500Å) long-slit spectrum of P Cygni obtained with the newly commissioned WINERED spectrograph in Japan. In the obtained spectrum, we have found that the velocity profiles of the [Fe  ii ] emission lines are resolved into two peaks at a velocity of ≃220 km s−1 with a moderate dip in between and with additional sub-peaks at ≃±100 km s−1. The sub-peak component is confirmed with the long-slit echellogram to originate in the known shell with a radius of ≃10 arcsec, which was originally created by the outburst in 1600 ad. On the other hand, the ≃220 km s−1 component, which dominates the [Fe  ii ] flux from P Cygni, is found to be concentrated closer to the central star with an apparent spatial extent of ≃3 arcsec. The extent is much larger than the compact (<0.1 arcsec) regions traced with hydrogen, helium, and metal permitted lines. The velocity, estimated mass, and dynamical time of the extended emission-line region suggest that the region is an outer part of the stellar wind region. We suggest that the newly identified emission-line region may trace a reverse shock due to the stellar wind overtaking the outburst shell. [ABSTRACT FROM AUTHOR]

Summary: MicroRNAs (miRNAs) are typically generated as ∼22-nucleotide double-stranded RNAs via the processing of precursor hairpins by the ribonuclease III enzyme Dicer, after which they are loaded into Argonaute (Ago) proteins to form an RNA-induced silencing complex (RISC). However, the biogenesis of miR-451, an erythropoietic miRNA conserved in vertebrates, occurs independently of Dicer and instead requires cleavage of the 3′ arm of the pre-miR-451 precursor hairpin by Ago2. The 3′ end of the Ago2-cleaved pre-miR-451 intermediate is then trimmed to the mature length by an unknown nuclease. Here, using a classical chromatographic approach, we identified poly(A)-specific ribonuclease (PARN) as the enzyme responsible for the 3′–5′ exonucleolytic trimming of Ago2-cleaved pre-miR-451. Surprisingly, our data show that trimming of Ago2-cleaved precursor miRNAs is not essential for target silencing, indicating that RISC is functional with miRNAs longer than the mature length. Our findings define the maturation step in the miRNA biogenesis pathway that depends on Ago2-mediated cleavage. [Copyright &y& Elsevier]

Heat shock protein 90 (Hsp90), a conserved molecular chaperone for a specific set of proteins critical for signal transduction including several oncogenic proteins, has been recognized as a promising target for anticancer therapy. Hsp90 inhibition also sensitizes cancer cells to DNA damage. However, the underlying mechanisms are not fully understood. Here, we provide evidence that Hsp90 is a general regulator of phosphatidylinositol 3-kinase-related protein kinase (PIKK) family proteins, central regulators of stress responses including DNA damage. Inhibition of Hsp90 causes a reduction of all PIKK and suppresses PIKK-mediated signaling. In addition, Hsp90 forms complexes with RUVBL1/2 complex and Tel2 complex, both of which have been shown to interact with all PIKK and control their abundance and functions. These results suggest that Hsp90 can form multiple complexes with the RUVBL1/2 complex and Tel2 complex and function in the regulation of PIKK, providing additional rationale for the effectiveness of Hsp90 inhibition for anticancer therapy, including sensitization to DNA damage. ( Cancer Sci 2012; 103: 50-57) [ABSTRACT FROM AUTHOR]

Proteins of the phosphatidylinositol 3-kinase-related protein kinase (PIKK) family are activated by various cellular stresses, including DNA damage, premature termination codon and nutritional status, and induce appropriate cellular responses. The importance of PIKK functions in the maintenance of genome integrity, accurate gene expression and the proper control of cell growth/proliferation is established. Recently, ATPase associated diverse cellular activities (AAA+) proteins RUVBL1 and RUVBL2 (RUVBL1/2) have been shown to be common regulators of PIKKs. The RUVBL1/2 complex regulates PIKK-mediated stress responses through physical interactions with PIKKs and by controlling PIKK mRNA levels. In this review, the functions of PIKKs in stress responses are outlined and the physiological significance of the integrated regulation of PIKKs by the RUVBL1/2 complex is presented. We also discuss a putative "PIKK regulatory chaperone complex" including other PIKK regulators, Hsp90 and the Tel2 complex [ABSTRACT FROM AUTHOR]

Drosophila melanogaster has two Dicer proteins with specialized functions. Dicer-1 liberates miRNA-miRNA* duplexes from precursor miRNAs (pre-miRNAs), whereas Dicer-2 processes long double-stranded RNAs into small interfering RNA duplexes. It was recently demonstrated that Dicer-2 is rendered highly specific for long double-stranded RNA substrates by inorganic phosphate and a partner protein R2D2. However, it remains unclear how Dicer-1 exclusively recognize pre-miRNAs. Here we show that fly Dicer-1 recognizes the single-stranded terminal loop structure of pre-miRNAs through its N-terminal helicase domain, checks the loop size and measures the distance between the 3? overhang and the terminal loop. This unique mechanism allows fly Dicer-1 to strictly inspect the authenticity of pre-miRNA structures. [ABSTRACT FROM AUTHOR]

We report the results of new survey of star-forming regions in the outer Galaxy at Galactocentric radius of more than 13.5 kpc, where the environment is significantly different from that in the solar neighborhood. [ABSTRACT FROM PUBLISHER]

Outskirts of spiral galaxies, including our own, and dwarf irregular galaxies are known to have a different environment from the solar neighborhood, e.g., low metallicities ( ~ − 1 dex). Among them, the outer Galaxy is the closest and hence is so far the only site suitable for population studies of resolved stars on the same basis as solar neighborhood. We have obtained NIR images of young clusters in the outer Galaxy, using the Subaru 8.2-m telescope, and clearly resolved cluster members with mass detection limits of ~ 0.1 M⊙. Based on the fitting of K-band luminosity functions (KLFs) for four clusters, we found that the initial mass function (IMF) in the outer Galaxy is consistent with that in the solar neighborhood in terms of the high-mass slope and IMF peak. Upcoming observations with a higher spatial resolution and sensitivity, using JWST, TMT, etc., will allow us to extend spatially-resolved studies of the IMF to Local Group galaxies. [ABSTRACT FROM PUBLISHER]

We present a comprehensive survey of diffuse interstellar bands (DIBs) in 0.91-1.32 μm with the newly developed near-infrared (NIR) spectrograph WINERED mounted on the Araki 1.3 m telescope in Japan. We obtained high-resolution (R = 28, 300) spectra of 25 early-type stars with color excesses of 0.07 < E(B – V) < 3.4. In addition to the 4 DIBs previously detected in this wavelength range, we identified 15 new DIBs, 7 of which were reported as DIB “candidates” by Cox. We analyze the correlations among NIR DIBs, strong optical DIBs, and the reddening of the stars. Consequently, we found that all NIR DIBs show weaker correlations with the reddening rather than the optical DIBs, suggesting that the equivalent widths of NIR DIBs depend on some physical conditions of the interstellar clouds, such as UV flux. Three NIR DIBs, λλ10780, 10792, and 11797, are found to be classifiable as a “family,” in which the DIBs are well correlated with each other, suggesting that the carriers of these DIBs are connected with some chemical reactions and/or have similar physical properties such as ionization potential. We also found that the three strongest NIR DIBs, λλ10780, 11797, and 13175 are well correlated with the optical DIB λ5780.5, whose carrier is proposed to be a cation molecule with high ionization potential, indicating that the carriers of the NIR DIBs could be cation molecules. [ABSTRACT FROM AUTHOR]

We report the discovery of star formation activity in perhaps the most distant molecular cloud in the extreme outer galaxy. We performed deep near-infrared imaging with the Subaru 8.2 m telescope, and found two young embedded clusters at two CO peaks of “Digel Cloud 1” at the kinematic distance of D = 16 kpc (Galactocentric radius RG = 22 kpc). We identified 18 and 45 cluster members in the two peaks, and the estimated stellar densities are ∼5 and ∼3 pc–2, respectively. The observed K-band luminosity function suggests that the age of the clusters is less than 1 Myr and also that the distance to the clusters is consistent with the kinematic distance. On the sky, Cloud 1 is located very close to the H I peak of high-velocity cloud Complex H, and there are some H I intermediate velocity structures between the Complex H and the Galactic disk, which could indicate an interaction between them. We suggest the possibility that Complex H impacting on the Galactic disk has triggered star formation in Cloud 1 as well as the formation of the Cloud 1 molecular cloud. [ABSTRACT FROM AUTHOR]

The basolateral tumor suppressor protein Lgl is important for the regulation of epithelial cell polarity and tissue morphology. Recent studies have shown a physical and functional interaction of Lgl with another polarity-regulating protein machinery, the apical PAR-3-aPKC-PAR-6 complex, in epithelial cells. However, the mechanism of Lgl-mediated regulation of epithelial cell polarity remains obscure. By an siRNA method, we here show that endogenous Lgl is required for the disassembly of apical membrane domains in depolarizing MDCK cells induced by Ca2+ depletion. Importantly, this Lgl function is mediated by the suppression of the apical PAR-3-aPKC-PAR-6 complex activity. Analysis using 2D- or 3D-cultured cells in collagen gel suggests the importance of this suppressive regulation of Lgl on the collagen-mediated re-establishment of apical membrane domains and lumen formation. These results indicate that basolateral Lgl plays a crucial role in the disassembly of apical membrane domains to induce the orientation of apical membrane polarity, which is mediated by the suppression of apical PAR3-aPKC-PAR-6 complex activity. [ABSTRACT FROM AUTHOR]