Control of Hsp90 chaperone and its clients by N-terminal acetylation and the N-end rule pathway

We found that the heat shock protein 90 (Hsp90) chaperone system of the yeast Saccharomyces cerevisiae is greatly impaired in naa10Δ cells, which lack the NatA N^α-terminal acetylase (Nt-acetylase) and therefore cannot N-terminally acetylate a majority of normally N-terminally acetylated proteins, including Hsp90 and most of its cochaperones. Chk1, a mitotic checkpoint kinase and a client of Hsp90, was degraded relatively slowly in wild-type cells but was rapidly destroyed in naa10Δ cells by the Arg/N-end rule pathway, which recognized a C terminus-proximal degron of Chk1. Diverse proteins (in addition to Chk1) that are shown here to be targeted for degradation by the Arg/N-end rule pathway in naa10Δ cells include Kar4, Tup1, Gpd1, Ste11, and also, remarkably, the main Hsp90 chaperone (Hsc82) itself. Protection of Chk1 by Hsp90 could be overridden not only by ablation of the NatA Nt-acetylase but also by overexpression of the Arg/N-end rule pathway in wild-type cells. Split ubiquitin-binding assays detected interactions between Hsp90 and Chk1 in wild-type cells but not in naa10Δ cells. These and related results revealed a major role of Nt-acetylation in the Hsp90-mediated protein homeostasis, a strong up-regulation of the Arg/N-end rule pathway in the absence of NatA, and showed that a number of Hsp90 clients are previously unknown substrates of the Arg/N-end rule pathway

Physical activity differences by birthplace and sex in youth of Mexican Heritage

Journal ArticleBackground: One of the goals of Healthy People 2020 is to increase physical activity (PA) and reduce health disparities among different racial and ethnic segments of the U.S. population. Few studies have been conducted to examine PA differences by birthplace and sex in youth of Mexican heritage. Methods: Participants were 101 youth (43 boys, 58 girls, 59% U.S.-born, 41% Mexico-born) who wore a sealed New Lifestyles NL-1000 pedometer for 7 consecutive days. Results: Mexico-born youth took more steps, on average, than their U.S.-born counterparts (P = .038). However, moderate-to-vigorous intensity (MVI) time did not differ between Mexico- and U.S.-born youth (P = .146). By contrast to birthplace, sex-related differences were more consistent, as boys took more steps (P = .005) and accumulated more MVI time (P = .043) than girls. Only 4% of our sample met either one or both of the PA recommendations. Conclusions: We conclude that PA may differ by birthplace and by sex in youth of Mexican heritage, as U.S.-born girls were the least active segment of our sample. Culturally sensitive interventions to increase daily PA must become a higher public health priority for youth of Mexican heritage, in particular, for U.S.-born girls of Mexican heritage

Five enzymes of the Arg/N-degron pathway form a targeting complex: The concept of superchanneling

The Arg/N-degron pathway targets proteins for degradation by recognizing their N-terminal (Nt) residues. If a substrate bears, for example, Nt-Asn, its targeting involves deamidation of Nt-Asn, arginylation of resulting Nt-Asp, binding of resulting (conjugated) Nt-Arg to the UBR1-RAD6 E3-E2 ubiquitin ligase, ligase-mediated synthesis of a substrate-linked polyubiquitin chain, its capture by the proteasome, and substrate’s degradation. We discovered that the human Nt-Asn–specific Nt-amidase NTAN1, Nt-Gln–specific Nt-amidase NTAQ1, arginyltransferase ATE1, and the ubiquitin ligase UBR1-UBE2A/B (or UBR2-UBE2A/B) form a complex in which NTAN1 Nt-amidase binds to NTAQ1, ATE1, and UBR1/UBR2. In addition, NTAQ1 Nt-amidase and ATE1 arginyltransferase also bind to UBR1/UBR2. In the yeast Saccharomyces cerevisiae, the Nt-amidase, arginyltransferase, and the double-E3 ubiquitin ligase UBR1-RAD6/UFD4-UBC4/5 are shown to form an analogous targeting complex. These complexes may enable substrate channeling, in which a substrate bearing, for example, Nt-Asn, would be captured by a complex-bound Nt-amidase, followed by sequential Nt modifications of the substrate and its polyubiquitylation at an internal Lys residue without substrate’s dissociation into the bulk solution. At least in yeast, the UBR1/UFD4 ubiquitin ligase interacts with the 26S proteasome, suggesting an even larger Arg/N-degron–targeting complex that contains the proteasome as well. In addition, specific features of protein-sized Arg/N-degron substrates, including their partly sequential and partly nonsequential enzymatic modifications, led us to a verifiable concept termed “superchanneling.” In superchanneling, the synthesis of a substrate-linked poly-Ub chain can occur not only after a substrate’s sequential Nt modifications, but also before them, through a skipping of either some or all of these modifications within a targeting complex

Comparisons of ELISA and Western blot assays for detection of autophagy flux

We analyzed autophagy/mitophagy flux in vitro (C2C12 myotubes) and in vivo (mouse skeletal muscle) following the treatments of autophagy inducers (starvation, rapamycin) and a mitophagy inducer (carbonyl cyanide m-chlorophenylhydrazone, CCCP) using two immunodetection methods, ELISA and Western blotting, and compared their working range, accuracy, and reliability. The ELISAs showed a broader working range than that of the LC3 Western blots (Table 1). Table 2 showed that data value distribution was tighter and the average standard error from the ELISA was much smaller than those of the Western blot, directly relating to the accuracy of the assay. Test-retest reliability analysis showed good reliability for three individual ELISAs (interclass correlation, ≥ 0.7), but poor reliability for three individual Western blots (interclass correlation, ≤ 0.4) (Table 3). Keywords: Autophagy, Mitophagy, ELISA, Western blot, Skeletal muscl

A Large-Scale 3D Face Mesh Video Dataset via Neural Re-parameterized Optimization

We propose NeuFace, a 3D face mesh pseudo annotation method on videos via neural re-parameterized optimization. Despite the huge progress in 3D face reconstruction methods, generating reliable 3D face labels for in-the-wild dynamic videos remains challenging. Using NeuFace optimization, we annotate the per-view/-frame accurate and consistent face meshes on large-scale face videos, called the NeuFace-dataset. We investigate how neural re-parameterization helps to reconstruct image-aligned facial details on 3D meshes via gradient analysis. By exploiting the naturalness and diversity of 3D faces in our dataset, we demonstrate the usefulness of our dataset for 3D face-related tasks: improving the reconstruction accuracy of an existing 3D face reconstruction model and learning 3D facial motion prior. Code and datasets will be available at https://neuface-dataset.github.io.Comment: 9 pages, 7 figures, and 3 tables for the main paper. 8 pages, 6 figures and 3 tables for the appendi

Ionothermal Synthesis of Metal-Organic Framework

Ionothermal synthesis employs ionic liquids for synthesis of metal organic frameworks (MOFs) as solvent and template. The cations and anions of ionic liquids may be finely adjusted to produce a great variety of reaction environments and thus frameworks. Organisation of the structures synthesised from related ionic liquid combinations give rise to provocative chemical trends that may be used to predict future outcomes. Further analysis of their structures is possible by reducing the complex framework to its underlying topology, which by itself brings more precision to prediction. Through reduction, many seemingly different, but related classes of structures may be merged into larger groups and provide better understanding of the nanoscopic structures and synthesis conditions that gave rise to them. Ionothermal synthesis has promised to enable us to effectively plan the synthesis ahead for a given purpose. However, for its promise to be kept, several difficult limitations must be overcome, including the inseparable cations from the solvent that reside in the framework pore

LaughTalk: Expressive 3D Talking Head Generation with Laughter

Laughter is a unique expression, essential to affirmative social interactions of humans. Although current 3D talking head generation methods produce convincing verbal articulations, they often fail to capture the vitality and subtleties of laughter and smiles despite their importance in social context. In this paper, we introduce a novel task to generate 3D talking heads capable of both articulate speech and authentic laughter. Our newly curated dataset comprises 2D laughing videos paired with pseudo-annotated and human-validated 3D FLAME parameters and vertices. Given our proposed dataset, we present a strong baseline with a two-stage training scheme: the model first learns to talk and then acquires the ability to express laughter. Extensive experiments demonstrate that our method performs favorably compared to existing approaches in both talking head generation and expressing laughter signals. We further explore potential applications on top of our proposed method for rigging realistic avatars.Comment: Accepted to WACV202

Selection of Elevation Models for Flood Inundation Map Generation in Small Urban Stream: Case Study of Anyang Stream

To reduce flood damages, the Ministry of Environment in Korea has provided a flood inundation map so that people can expediently identify flood-prone areas. However, the current flood inundation maps have been produced based on the DEM which makes it difficult to represent realistic situations due to the lack of reproduction of land surface conditions. This study aims to provide more accurate and detailed flood inundation maps for flooding events due to river overflow in small urban areas. In this study, flood inundation analysis is performed using the river analysis system, HEC-RAS 2D, with the DSM and the DEM of urban areas in the Anyang Stream Basin, Korea to examine the differences in terms of terrain data and flooded area. Finally, for urban areas with dense buildings and congested road networks, the flood inundation analysis based on DSM can represent a more realistic flood situation and create an appropriate flood inundation map