Dissolution of ribonucleoprotein condensates by the embryonic stem cell protein L1TD1.

L1TD1 is a cytoplasmic RNA-binding protein specifically expressed in pluripotent stem cells and, unlike its mouse ortholog, is essential for the maintenance of stemness in human cells. Although L1TD1 is the only known protein-coding gene domesticated from a LINE-1 (L1) retroelement, the functional legacy of its ancestral protein, ORF1p of L1, and how it is manifested in L1TD1 are still unknown. Here, we determined RNAs associated with L1TD1 and found that, like ORF1p, L1TD1 binds L1 RNAs and localizes to high-density ribonucleoprotein (RNP) condensates. Unexpectedly, L1TD1 enhanced the translation of a subset of mRNAs enriched in the condensates. L1TD1 depletion promoted the formation of stress granules in embryonic stem cells. In HeLa cells, ectopically expressed L1TD1 facilitated the dissolution of stress granules and granules formed by pathological mutations of TDP-43 and FUS. The glutamate-rich domain and the ORF1-homology domain of L1TD1 facilitated dispersal of the RNPs and induced autophagy, respectively. These results provide insights into how L1TD1 regulates gene expression in pluripotent stem cells. We propose that the ability of L1TD1 to dissolve stress granules may provide novel opportunities for treatment of neurodegenerative diseases caused by disturbed stress granule dynamics.

Global analysis of binding sites of U2AF1 and ZRSR2 reveals RNA elements required for mutually exclusive splicing by the U2- and U12-type spliceosome.

Recurring mutations in genes encoding 3' splice-site recognition proteins, U2AF1 and ZRSR2 are associated with human cancers. Here, we determined binding sites of the proteins to reveal that U2-type and U12-type splice sites are recognized by U2AF1 and ZRSR2, respectively. However, some sites are spliced by both the U2-type and U12-type spliceosomes, indicating that well-conserved consensus motifs in some U12-type introns could be recognized by the U2-type spliceosome. Nucleotides flanking splice sites of U12-type introns are different from those flanking U2-type introns. Remarkably, the AG dinucleotide at the positions -1 and -2 of 5' splice sites of U12-type introns with GT-AG termini is not present. AG next to 5' splice site introduced by a single nucleotide substitution at the -2 position could convert a U12-type splice site to a U2-type site. The class switch of introns by a single mutation and the bias against G at the -1 position of U12-type 5' splice site support the notion that the identities of nucleotides in exonic regions adjacent to splice sites are fine-tuned to avoid recognition by the U2-type spliceosome. These findings may shed light on the mechanism of selectivity in U12-type intron splicing and the mutations that affect splicing.

Toxoplasma gondii and Rickettsia spp. in ticks collected from migratory birds in the Republic of Korea.

Migratory birds disperse ticks and associated tick-borne pathogens along their migratory routes. Four selected pathogens of medical importance (Coxiella burnetii, Rickettsia spp., Francisella tularensis, and Toxoplasma gondii) were targeted for detection in 804 ticks (365 pools) collected from migratory birds at Hong and Heuksan Islands in the Republic of Korea (ROK) from 2010 to 2011 and 2016. Toxoplasma gondii and Rickettsia spp., were detected in 1/365 (0.27%) and 34/365 (9.32%) pools of ticks, respectively. T. gondii and five rickettsial species were recorded in ticks collected from migratory birds for the first time in ROK. The five rickettsial species (R. monacensis, Candidatus Rickettsia longicornii, R. japonica, R. raoultii, and R. tamurae) were identified using sequence and phylogenetic analysis using ompA and gltA gene fragments. Rickettsia spp. are important pathogens that cause rickettsiosis in humans, with cases recorded in the ROK. These results provide important evidence for the potential role of migratory birds in the introduction and dispersal of T. gondii and Rickettsia spp. along their migratory routes and raise awareness of potential transmission of zoonotic tick-borne pathogens associated with migratory birds in the ROK.

Plastic debris ingestion by seabirds on the Korean Peninsula.

Plastic ingestion studies in seabirds that analyse the frequency of occurrence and the characteristics of the plastics ingested by each species provide valuable information for marine environmental assessments by quantifying the impacts of marine debris on seabirds. We investigated the frequency of plastic ingestion from a sample of 387 individuals of 11 seabird species on the Korean Peninsula. We found evidence of plastic ingestion in red-breasted mergansers (Mergus serrator) (33.3%), Pacific loons (Gavia pacifica) (10.0%), Swinhoe's storm petrels (Hydrobates monorhis) (93.7%), black-tailed gulls (Larus crassirostris) (12.9%) and ancient murrelets (Synthliboramphus antiquus) (0.9%). In particular, it was observed that Swinhoe's storm petrels had the highest frequency of plastic ingestion, both in terms of the number of affected individuals, and the accumulated mass of plastic debris ingested. The majority of seabirds examined in our study had ingested microplastics, comprised predominantly of user plastics. This is the first report quantifying plastic ingestion in seabirds on the Korean Peninsula and in the broader area of the East Asian Seas.

Molecular Detection and Phylogenetic Analysis of Anaplasma and Borrelia Species in Ticks Collected from Migratory Birds at Heuksan, Hong, and Nan Islands, Republic of Korea.

The extended distribution and potential introduction of exotic ticks and associated tick-borne pathogens along the northern and southern routes of migratory birds pose zoonotic tick-borne disease risks to wild and domestic animals and incidentally to humans. A knowledge of bird migratory patterns, species of attached ticks, and associated pathogens during their migrations to and from their feeding and nesting grounds is central to understanding associated tick-borne disease risks. Tick-borne disease surveillance was conducted from 2010 to 2011 and 2016 at Hong-do (do = island), Heuksan-do, and Nan-do, major stopovers for migratory birds in Republic of Korea (ROK), as part of the Migratory Birds Research Center bird-banding program for studying bird migration patterns in the ROK. A total of 877 ticks belonging to three genera and nine species were collected, Ixodes turdus (576, 65.7%), Haemaphysalis flava (134, 15.3%), H. longicornis (91, 10.4%), I. nipponensis (56, 6.4%), H. formosensis (7, 0.8%), H. ornithophila (6, 0.7%), H. phasiana (5, 0.6%), H. concinna (1, 0.1%), and Amblyomma testudinarium (1, 0.1%) were collected from 274 birds belonging to 20 genera and 41 species. A total of 15/380 pools (3.95%) were positive for Borrelia species (14 pools of I. turdus and 1 pool of H. flava), while only 1/380 pools (0.26%) was positive for Anaplasma phagocytophilum (1 pool of I. nipponensis). Our findings support the role of migratory birds as possible vectors for the introduction of tick-borne pathogens, which requires continuous monitoring for the potential introduction of ticks and their associated tick-borne pathogens.

The conserved microRNA miR-8-3p coordinates the expression of V-ATPase subunits to regulate ecdysone biosynthesis for Drosophila metamorphosis.

The steroid hormone ecdysone is the central regulator of insect metamorphosis, during which a growing, immature larva is remodeled, through pupal stages, to a reproductive adult. However, the underlying mechanisms of ecdysone-mediated metamorphosis remain to be fully elucidated. Here, we identified metamorphosis-associated microRNAs (miRNAs) and their potential targets by cross-linking immunoprecipitation coupled with deep sequencing of endogenous Argonaute 1 protein in Drosophila. Interestingly, miR-8-3p targeted five Vha genes encoding distinct subunits of vacuolar H+ -ATPase (V-ATPase), which has a vital role in the organellar acidification. The expression of ecdysone-responsive miR-8-3p is normally downregulated during Drosophila metamorphosis, but temporary overexpression of miR-8-3p in the whole body at the end of larval development led to defects in metamorphosis and survival, hallmarks of aberrant ecdysone signaling. In addition, miR-8-3p was expressed in the prothoracic gland (PG), which produces and releases ecdysone in response to prothoracicotropic hormone (PTTH). Notably, overexpression of miR-8-3p or knockdown of its Vha targets in the PG resulted in larger than normal, ecdysone-deficient larvae that failed to develop into the pupal stage but could be rescued by ecdysone feeding. Moreover, these animals showed defective PTTH signaling with a concomitant decrease in the expression of ecdysone biosynthetic genes. We also demonstrated that the regulatory network between the conserved miR-8-3p/miR-200 family and V-ATPase was functional in human cells. Consequently, our data indicate that the coordinated regulation of V-ATPase subunits by miR-8-3p is involved in Drosophila metamorphosis by controlling the ecdysone biosynthesis.

MicroRNAs of miR-17-92 cluster increase gene expression by targeting mRNA-destabilization pathways.

MicroRNAs (miRNAs) of the miR-17-92 cluster are overexpressed in human cancers, and their enforced expression is tumorigenic in mouse models. A number of genes are reported to be targets of these miRNAs and are implicated in their tumorigenic potential. However, the mode of action by miRNAs suggests that global analysis of their targets is required to understand their cellular roles. In this study, we globally analyzed AGO2-bound mRNAs and found that the miR-17-92 miRNAs coherently repress multiple targets involved in the destabilization of mRNA. While the miRNAs repress the expression of their targets, they increase stability and lengthen the poly-A tails of non-target mRNAs. Furthermore, the expression of BTG3, TOB1, CSNK1A1 and ANKRD52 is negatively correlated with the expression of the miR-17-92 cluster in cancer cell lines. Our results suggest that the miR-17-92 miRNAs promote tumorigenesis not only by repression of key regulators, but also by posttranscriptional increases of global gene expression.

Ecdysone-responsive microRNA-252-5p controls the cell cycle by targeting Abi in Drosophila.

The steroid hormone ecdysone has a central role in the developmental transitions of insects through its control of responsive protein-coding and microRNA (miRNA) gene expression. However, the complete regulatory network controlling the expression of these genes remains to be elucidated. In this study, we performed cross-linking immunoprecipitation coupled with deep sequencing of endogenous Argonaute 1 (Ago1) protein, the core effector of the miRNA pathway, in Drosophila S2 cells. We found that regulatory interactions between miRNAs and their cognate targets were substantially altered by Ago1 in response to ecdysone signaling. Additionally, during the larva-to-adult metamorphosis, miR-252-5p was up-regulated via the canonical ecdysone-signaling pathway. Moreover, we provide evidence that miR-252-5p targets Abelson interacting protein ( Abi) to decrease the protein levels of cyclins A and B, controlling the cell cycle. Overall, our data suggest a potential role for the ecdysone/miR-252-5p/Abi regulatory axis partly in cell-cycle control during metamorphosis in Drosophila.-Lim, D.-H., Lee, S., Han, J. Y., Choi, M.-S., Hong, J.-S., Seong, Y., Kwon, Y.-S., Lee, Y. S. Ecdysone-responsive microR-252-5p controls the cell cycle by targeting Abi in Drosophila.

Global analysis of AGO2-bound RNAs reveals that miRNAs induce cleavage of target RNAs with limited complementarity.

Among the four Argonaute family members in mammals, only AGO2 protein retains endonuclease activity and facilitates cleavage of target RNAs base-pairing with highly complementary guide RNAs. Despite the deeply conserved catalytic activity, only a small number of targets have been reported to extensively base pair with cognate miRNAs to be cleaved by AGO2. Here, we analyzed AGO2-bound RNAs by CrossLinking ImmunoPrecipitation (CLIP) of genetically modified cells that express epitope-tagged AGO2 from the native genomic locus. We found that HMGA2 mRNA is cleaved by AGO2 loaded with let-7 and miR-21. In contrast to the generally accepted notion, the base-pairing from the seed region to the cleavage site, rather than perfect or near perfect complementarity, was required for cleavage of the target mRNA in cells. Non-templated addition of nucleotides at the 3' end of the cleaved RNA was observed, further supporting the AGO2-mediated cleavage. Based on the observation that the limited complementarity is the minimum requirement for cleavage, we found that AGO2-mediated cleavage of targets is more common than previously thought. Our result may explain the vital role of endonuclease activity in controlling miRNA-mediated gene regulation.

Characteristics of Intramolecular Charge Transfer by J-Aggregates in Merocyanine Dye LB Films.

In this study, for the development of future molecular electronic devices, we have investigated the characteristics of the aggregates of Langmuir-Blodgett films. The characteristics of intramolecular charge transfer by J-aggregates in merocyanine dye LB films have been studied experimentally by using UV irradiation and heat treatment. In addition to intramolecular charge transfer, we also studied the conjugation and energy changes of the molecules. In case a dye is thinned by LB method, the alkyl chain is often displaced in order to form a mono-molecular film with ease. Since the molecular association form is often made by self-organization of molecules themselves, in case the dye and the alkyl chain are strongly bonded by the covalent bond, it may be said that the properties of the LB film to be built up are almost determined at the time of synthesis of film-forming molecules. Meanwhile, since, in case LB film is fabricated by the diffusion absorption method, the cohesive force between the water-soluble dye and the surface-active mono-molecular film is electrostatic, the dye molecule can move relatively freely on the air/water interface, which may be regarded as a two-dimensional crystal growth process.

White OLED in Hybrid Structure for Enhancing Color Purity.

We synthesized the red emission material, bis(1,4-bis(5-phenyloxazol-2-yl)phenyl) iridium(picolate) [Ir-complexes] and the blue emission material, bis (2-(2-hydroxyphenyl) benzoxazolate)zinc [Zn(HPB)2]. White Organic Light Emitting Diodes were fabricated by using Zn(HPB)2 for a blue emitting layer, Ir-complexes for a red emitting layer and a tris (8-hydroxy quinoline)aluminum [Alq3] for a green emitting layer. The important experimental results obtained, white OLED was fabricated by using double emitting layers of Zn(HPB)2 and Alq3:Ir-complexes, and hole blocking layer of 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline[BCP]. We also varied the thickness of BCP. When the thickness of BCP layer was 5 nm, white emission was achieved. We obtained a maximum luminance of 5400 cd/m2 at a current density of 650 mA/cm2. The CIE coordinates was (0.339, 0.323) at voltage of 10 V.

Hybrid Structure White Organic Light Emitting Diode for Enhanced Efficiency by Varied Doping Rate.

Novel materials based on Zn(HPB)2 and Ir-complexes were synthesized as blue or red emitters, respectively. White organic light emitting diodes were fabricated using the Zn(HPB)2 as a blue emitting layer, Ir-complexes as a red emitting layer and Alq3 as a green emitting layer. The obtained experimental results, were based on white OLEDs fabricated using double emission layers of Zn(HPB)2 and Alq3:Ir-complexes. The doping rate of the Ir-complexes was varied at 0.4%, 0.6%, 0.8% and 1.0%. When the doping rate of the Alq3:Ir-complexes was 0.6%, a white emission was achieved. The Commission Internationale de l'Eclairage coordinates of the device's white emission were (0.316, 0.331) at an applied voltage of 10.75 V.

Ornithodoros sawaii (Ixodida: Argasidae) Larvae Collected from Hydrobates monorhis on Sogugul and Gaerin Islands, Jeollanam-do (Province), Republic of Korea.

The 65th Medical Brigade and Public Health Command District-Korea, in collaboration with the Migratory Bird Research Center, National Park Research Institute, conducted migratory bird tick surveillance at Sogugul and Gaerin Islands (small rocky bird nesting sites), Jeollanam-do (Province), Republic of Korea (ROK), on 30 July and 1 August 2009. Breeding seabirds captured by hands in their nesting burrows were banded, identified to species, and carefully examined for ticks during the nesting season. A total of 9 Ornithodoros sawaii larvae were removed from 4 adult Hydrobates monorhis (Swinhoe's storm petrel). The identification of the larvae of O. sawaii collected from migratory seabirds were molecularly confirmed using mitochondrial 16S rDNA primer sets.

A Study on the Rectification Property of Self-Assembled Viologen Single Molecules Using a Scanning Tunneling Microscopy.

An ultrahigh vacuum scanning tunneling microscopy (UHV-STM) and a scanning tunneling spectroscopy (STS) are used measure the rectification property of self-assembled viologen single molecules (VC8SH, VC10SH, HSC8VC8SH, and HSC10VC10SH) in the previous study. Using STM we observe viologen single molecules in the self-assembled octanethiol (OT) SAM matrix. In the OT matrix a mixed phase that includes a c(4 x 2) superlattice of high-density standing up-phase is observed. We indicate high peak current-like rectifications at + 1.68 V(VC8SH), + 1.56 V(VC10SH), + 1.14 V(HSC8VC8SH), and + 1.04 V(HSC10VC10SH) based on the experiment implemented in this study. In addition, transition voltages (Vtrans) from direct tunneling to the Fowler-Nordheim tunneling are presented at 1.08 V(VC8SH), 0.97 V(VC10SH), 0.99 V(HSC8VC8SH), and 0.89 V(HSC1VC1SH).

Fabrication of White Organic Light Emitting Diode Using Two Types of Zn-Complexes as an Emitting Layer.

We have studied white OLED using two types of Zn-complexes as an emitting layer. We synthesized brand new two emissive materials, Zn(HPQ)2 as a yellow emitting material and Zn(HPB)2 as a blue emitting material. The Zn-complexes are low-molecular compounds and stable thermally. The fundamental structures of the fabricated OLED was ITO/NPB (40 nm)/Zn(HPB)2 (30 nm)/Zn(HPQ)2/LiF/Al. We varied the thickness of the Zn(HPQ)2 layer by 20, 30, and 40 nm. When the thickness of the Zn(HPQ)2 layer was 20 nm, the white emission was achieved. The maximum luminance was 12,000 cd/m2 at a current density of 800 mA/cm2. The CIE coordinates of the white emission were (0.319, 0.338) at an applied voltage of 10 V.

Spatial patterns, ecological niches, and interspecific competition of avian brood parasites: inferring from a case study of Korea.

Since obligate avian brood parasites depend completely on the effort of other host species for rearing their progeny, the availability of hosts will be a critical resource for their life history. Circumstantial evidence suggests that intense competition for host species may exist not only within but also between species. So far, however, few studies have demonstrated whether the interspecific competition really occurs in the system of avian brood parasitism and how the nature of brood parasitism is related to their niche evolution. Using the occurrence data of five avian brood parasites from two sources of nationwide bird surveys in South Korea and publically available environmental/climatic data, we identified their distribution patterns and ecological niches, and applied species distribution modeling to infer the effect of interspecific competition on their spatial distribution. We found that the distribution patterns of five avian brood parasites could be characterized by altitude and climatic conditions, but overall their spatial ranges and ecological niches extensively overlapped with each other. We also found that the predicted distribution areas of each species were generally comparable to the realized distribution areas, and the numbers of individuals in areas where multiple species were predicted to coexist showed positive relationships among species. In conclusion, despite following different coevolutionary trajectories to adapt to their respect host species, five species of avian brood parasites breeding in South Korea occupied broadly similar ecological niches, implying that they tend to conserve ancestral preferences for ecological conditions. Furthermore, our results indicated that contrary to expectation interspecific competition for host availability between avian brood parasites seemed to be trivial, and thus, play little role in shaping their spatial distributions and ecological niches. Future studies, including the complete ranges of avian brood parasites and ecological niches of host species, will be worthwhile to further elucidate these issues.

Global identification of target recognition and cleavage by the Microprocessor in human ES cells.

The Microprocessor plays an essential role in canonical miRNA biogenesis by facilitating cleavage of stem-loop structures in primary transcripts to yield pre-miRNAs. Although miRNA biogenesis has been extensively studied through biochemical and molecular genetic approaches, it has yet to be addressed to what extent the current miRNA biogenesis models hold true in intact cells. To address the issues of in vivo recognition and cleavage by the Microprocessor, we investigate RNAs that are associated with DGCR8 and Drosha by using immunoprecipitation coupled with next-generation sequencing. Here, we present global protein-RNA interactions with unprecedented sensitivity and specificity. Our data indicate that precursors of canonical miRNAs and miRNA-like hairpins are the major substrates of the Microprocessor. As a result of specific enrichment of nascent cleavage products, we are able to pinpoint the Microprocessor-mediated cleavage sites per se at single-nucleotide resolution. Unexpectedly, a 2-nt 3' overhang invariably exists at the ends of cleaved bases instead of nascent pre-miRNAs. Besides canonical miRNA precursors, we find that two novel miRNA-like structures embedded in mRNAs are cleaved to yield pre-miRNA-like hairpins, uncoupled from miRNA maturation. Our data provide a framework for in vivo Microprocessor-mediated cleavage and a foundation for experimental and computational studies on miRNA biogenesis in living cells.

SRSF2 promotes splicing and transcription of exon 11 included isoform in Ron proto-oncogene.

The product of proto-oncogene Ron is a human receptor for the macrophage-stimulating protein (MSP). Upon activation, Ron is able to induce cell dissociation, migration and matrix invasion. Exon 11 skipping of Ron pre-mRNA produces Ron△165 protein that is constitutively active even in the absence of its ligand. Here we show that knockdown of SRSF2 promotes the decrease of exon 11 inclusion, whereas overexpression of SRSF2 promotes exon 11 inclusion. We demonstrate that SRSF2 promotes exon 11 inclusion through splicing and transcription procedure. We also present evidence that reduced expression of SRSF2 induces a decrease in the splicing of both introns 10 and 11; by contrast, overexpression of SRSF2 induces an increase in the splicing of introns 10 and 11. Through mutation analysis, we show that SRSF2 functionally targets and physically interacts with CGAG sequence on exon 11. In addition, we reveal that the weak strength of splice sites of exon 11 is not required for the function of SRSF2 on the splicing of Ron exon 11. Our results indicate that SRSF2 promotes exon 11 inclusion of Ron proto-oncogene through targeting exon 11. Our study provides a novel mechanism by which Ron is expressed.

Well-defined all-conducting block copolymer bilayer hybrid nanostructure: selective positioning of lithium ions and efficient charge collection.

A block copolymerization of nonfunctionalized conducting monomers was developed to enable the successful synthesis of a highly insoluble 3,4-(ethylenedioxy)thienyl-based all-conducting block copolymer (PEDOT-b-PEDOT-TB) that could encapsulate nanocrystalline dyed TiO2 particles, resulting in the formation of an all-conducting block copolymer bilayer hybrid nanostructure (TiO2/Dye/PEDOT-b-PEDOT-TB). Lithium ions were selectively positioned on the outer PEDOT-TB surface. The distances through which the positively charged dye and PEDOT-TB(Li(+)) interacted physically or through which the TiO2 electrode and the Li(+) centers on PEDOT-TB(Li(+)) interacted ionically were precisely tuned and optimized within ca. 1 nm by controlling the thickness of the PEDOT blocking layer (the block length). The optimized structure provided efficient charge collection in an iodine-free dye-sensitized solar cell (DSC) due to negligible recombination of photoinduced electrons with cationic species and rapid charge transport, which improved the photovoltaic performance (η = 2.1 → 6.5%).

White organic light-emitting diodes with Zn-complexes.

This paper reviews OLEDs fabricated using Zn-complexes. Zn(HPB)2, Zn(HPB)q, and Zn(phen)q were synthesized as new electroluminescence materials. The electron affinity (EA) and ionization potential (IP) of Zn complexes were also determined and devices were characterized. Zn complexes such as Zn(HPB)2, Zn(HPB)q, and Zn(phen)q were found to exhibit blue and yellow emissions with wavelengths of 455, 532, and 535 nm, respectively. On the other hand, Zn(HPB)2 and Zn(HPB)q were applied as hole-blocking materials. As a result, the OLED efficiency by using Zn(HPB)2 as a hole-blocking material was improved. In particular, the OLED property of Zn(HPB)2 was found to be better than that of Zn(HPB)q. Moreover, Zn(phen)q was used as an electron-transporting material and compared with Alq3. The performance of the device with Zn(phen)q as an electron-transporting material was improved compared with Alq3-based devices. The Zn complexes can possibly be used as hole-blocking and electron-transporting materials in OLED devices. A white emission was ultimately realized from the OLED devices using Zn-complexes as inter-layer components.