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1.
Nat Commun ; 12(1): 5057, 2021 08 20.
Article in English | MEDLINE | ID: mdl-34417449

ABSTRACT

Argonaute is the primary mediator of metazoan miRNA targeting (MT). Among the currently identified >1,500 human RNA-binding proteins (RBPs), there are only a handful of RBPs known to enhance MT and several others reported to suppress MT, leaving the global impact of RBPs on MT elusive. In this study, we have systematically analyzed transcriptome-wide binding sites for 150 human RBPs and evaluated the quantitative effect of individual RBPs on MT efficacy. In contrast to previous studies, we show that most RBPs significantly affect MT and that all of those MT-regulating RBPs function as MT enhancers rather than suppressors, by making the local secondary structure of the target site accessible to Argonaute. Our findings illuminate the unappreciated regulatory impact of human RBPs on MT, and as these RBPs may play key roles in the gene regulatory network governed by metazoan miRNAs, MT should be understood in the context of co-regulating RBPs.


Subject(s)
MicroRNAs/metabolism , RNA-Binding Proteins/metabolism , 3' Untranslated Regions/genetics , Binding Sites , Evolution, Molecular , HeLa Cells , Hep G2 Cells , Humans , MicroRNAs/genetics , Nucleic Acid Conformation , Protein Binding , RNA, Messenger/chemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reproducibility of Results , Substrate Specificity
2.
Nat Commun ; 12(1): 5120, 2021 08 25.
Article in English | MEDLINE | ID: mdl-34433827

ABSTRACT

COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which infected >200 million people resulting in >4 million deaths. However, temporal landscape of the SARS-CoV-2 translatome and its impact on the human genome remain unexplored. Here, we report a high-resolution atlas of the translatome and transcriptome of SARS-CoV-2 for various time points after infecting human cells. Intriguingly, substantial amount of SARS-CoV-2 translation initiates at a novel translation initiation site (TIS) located in the leader sequence, termed TIS-L. Since TIS-L is included in all the genomic and subgenomic RNAs, the SARS-CoV-2 translatome may be regulated by a sophisticated interplay between TIS-L and downstream TISs. TIS-L functions as a strong translation enhancer for ORF S, and as translation suppressors for most of the other ORFs. Our global temporal atlas provides compelling insight into unique regulation of the SARS-CoV-2 translatome and helps comprehensively evaluate its impact on the human genome.


Subject(s)
COVID-19/virology , Protein Biosynthesis , SARS-CoV-2/genetics , Transcriptome , Gene Expression Regulation, Viral , Genome, Human , Humans , Open Reading Frames , RNA, Viral/genetics , RNA, Viral/metabolism , SARS-CoV-2/metabolism , Viral Proteins/genetics , Viral Proteins/metabolism
3.
Nanoscale ; 8(7): 4063-9, 2016 Feb 21.
Article in English | MEDLINE | ID: mdl-26819189

ABSTRACT

We investigated the subdomain structures of single-layer graphene oxide (GO) by characterizing local friction and conductance using conductive atomic force microscopy. Friction and conductance mapping showed that a single-layer GO flake has subdomains several tens to a few hundreds of nanometers in lateral size. The GO subdomains exhibited low friction (high conductance) in the sp(2)-rich phase and high friction (low conductance) in the sp(3)-rich phase. Current-voltage spectroscopy revealed that the local current flow in single-layer GO depends on the quantity of hydroxyl and carboxyl groups, and epoxy bridges within the 2-dimensional carbon layer. The presence of subdomains with different sp(2)/sp(3) carbon ratios on a GO flake was also confirmed by chemical mapping using scanning transmission X-ray microscopy. These results suggest that spatial mapping of the friction and conductance can be used to rapidly identify the composition of heterogeneous single-layer GO at nanometer scale, which is essential for understanding charge transport in nanoelectronic devices.

4.
ACS Appl Mater Interfaces ; 8(3): 1943-50, 2016 Jan 27.
Article in English | MEDLINE | ID: mdl-26734845

ABSTRACT

This study reports a method for the facile and high-yield exfoliation of WX2 (X = S, Se) by sonication under aqueous conditions using single-stranded DNA (abbreviated as ssDNA) of high molecular weight. The ssDNA provided a high degree of stabilization and prevented reaggregation, and it enhanced the exfoliation efficiency of WX2 nanosheets due to adsorption on the WX2 surface and the electrostatic repulsion of sugars in the ssDNA backbone. The exfoliation yield was higher with ssDNA (80%-90%) than without (2%-4%); the yield with ssDNA was also higher than the value previously reported for aqueous exfoliation (∼10%). Given that two-dimensional nanomaterials have potential health and environmental applications, we investigated antibacterial activity of exfoliated WX2-ssDNA nanosheets, relative to graphene oxide (GO), and found that WSe2-ssDNA nanosheets had higher antibacterial activity against Escherichia coli K-12 MG1655 cells than GO. Our method enables large-scale exfoliation in an aqueous environment in a single step with a short reaction time and under ambient conditions, and it can be used to produce surface-active or catalytic materials that have broad applications in biomedicine and other areas.


Subject(s)
Anti-Bacterial Agents/pharmacology , Chalcogens/pharmacology , DNA/chemistry , Tungsten/pharmacology , Escherichia coli/drug effects , Graphite , Hydrodynamics , Microbial Sensitivity Tests , Particle Size , Photoelectron Spectroscopy , Spectrophotometry, Ultraviolet , Spectrum Analysis, Raman
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