A Method to Generate and Analyze Modified Myristoylated Proteins

Covalent lipid modification of proteins is essential to their cellular localizations and functions. Engineered lipid motifs, coupled with bio-orthogonal chemistry, have been utilized to identify myristoylated or palmitoylated proteins in cells. However, whether modified proteins have similar properties as endogenous ones has not been well investigated mainly due to lack of methods to generate and analyze purified proteins. We have developed a method that utilizes metabolic interference and mass spectrometry to produce and analyze modified, myristoylated small GTPase ADP-ribosylation factor 1 (Arf1). The capacities of these recombinant proteins to bind liposomes and load and hydrolyze GTP were measured and compared with the unmodified myristoylated Arf1. The ketone-modified myristoylated Arf1 could be further labeled by fluorophore-coupled hydrazine and subsequently visualized through fluorescence imaging. This methodology provides an effective model system to characterize lipid-modified proteins with additional functions before applying them to cellular systems

Improved Differential Cryptanalysis on SPECK Using Plaintext Structures

Plaintext structures are a commonly-used technique for improving differential cryptanalysis. Generally, there are two types of plaintext structures: multiple-differential structures and truncated-differential structures. Both types have been widely used in cryptanalysis of S-box-based ciphers while for SPECK, an Addition-Rotation-XOR (ARX) cipher, the truncated-differential structure has not been used so far. In this paper, we investigate the properties of modular addition and propose a method to construct truncated-differential structures for SPECK. Moreover, we show that a combination of both types of structures is also possible for SPECK. For recovering the key of SPECK, we propose dedicated algorithms and apply them to various differential distinguishers, which helps to obtain a series of improved attacks on all variants of SPECK. Notably, on SPECK128, the time complexity of the attack can be reduced by a factor up to 2^15. The results show that the combination of both structures helps to improve the data and time complexity at the same time, as in the cryptanalysis of S-box-based ciphers

Structure–activity relationship studies of QS11, a small molecule Wnt synergistic agonist

Both the Wnt/β-catenin signaling pathway and small GTPases of the ADP-ribosylation factors (ARF) family play important roles in regulating cell development, homeostasis and fate. The previous report of QS11, a small molecule Wnt synergist that binds to ARF GTPase-activating protein 1 (ARFGAP1), suggests a role for ARFGAP1 in the Wnt/β-catenin pathway. However, direct inhibition of enzymatic activity of ARFGAP1 by QS11 has not been established. Whether ARFGAP1 is the only target that contributes to QS11's Wnt synergy is also not clear. Here we present structure-activity relationship (SAR) studies of QS11 analogs in two assays: direct inhibition of enzymatic activity of purified ARFGAP1 protein and cellular activation of the Wnt/β-catenin pathway. The results confirm the direct inhibition of ARFGAP1 by QS11, and also suggest the presence of other potential cellular targets of QS11

The genome of hibiscus hamabo reveals its adaptation to saline and waterlogged habitat

Hibiscus hamabo is a semi-mangrove species with strong tolerance to salt and waterlogging stress. However, the molecular basis and mechanisms that underlie this strong adaptability to harsh environments remain poorly understood. Here, we assembled a high-quality, chromosome-level genome of this semi-mangrove plant and analyzed its transcriptome under different stress treatments to reveal regulatory responses and mechanisms. Our analyses suggested that H. hamabo has undergone two recent successive polyploidy events, a whole-genome duplication followed by a whole-genome triplication, resulting in an unusually large gene number (107 309 genes). Comparison of the H. hamabo genome with that of its close relative Hibiscus cannabinus, which has not experienced a recent WGT, indicated that genes associated with high stress resistance have been preferentially preserved in the H. hamabo genome, suggesting an underlying association between polyploidy and stronger stress resistance. Transcriptomic data indicated that genes in the roots and leaves responded differently to stress. In roots, genes that regulate ion channels involved in biosynthetic and metabolic processes responded quickly to adjust the ion concentration and provide metabolic products to protect root cells, whereas no such rapid response was observed from genes in leaves. Using co-expression networks, potential stress resistance genes were identified for use in future functional investigations. The genome sequence, along with several transcriptome datasets, provide insights into genome evolution and the mechanism of salt and waterlogging tolerance in H. hamabo, suggesting the importance of polyploidization for environmental adaptation.DATA AVAILABILITY: The data supporting the findings of this work are available within the paper and its Supporting Information files. The data sets generated and analyzed during this study are available from the corresponding author upon request. All the whole-genome raw data generated during this study have been deposited in the SRA database under BioProject number PRJNA759075. Transcriptome clean data have been deposited in the SRA database under BioProject number PRJNA759717. The final chromosome-scale genome assembly and annotation data have been deposited in the Figshare database (https://doi.org/10.6084/m9.figshare.19142558.v1).Six Talent Peaks Project of Jiangsu Province (NY-042); Open Fund of the Jiangsu Key Laboratory for the Research and Utilization of Plant Resources (JSPKLB201928); Talent Training Funds of the Institute of Botany, Jiangsu Province and Chinese Academy of Sciences.https://academic.oup.com/hrBiochemistryGeneticsMicrobiology and Plant Patholog

Research on the action characteristics and breaking control of 100 kA series double break DC vacuum circuit breaker for CRAFT quench protection system

The DC vacuum circuit breaker plays an important role in the quench protection system of the Comprehensive Research Facility for Fusion Technology (CRAFT) project. The study of the operating characteristics of vacuum circuit breakers can effectively improve breaking reliability. This article introduces the design and basis of the series double break DC vacuum circuit breaker for the CRAFT quench protection system. It adopts the electromagnetic repulsion driving method to ensure rapid action and the operating mechanism design to ensure the synchronization of the double break action. A magnetic shielding setting is proposed through simulation calculations to solve the uniformity of magnetic induction strength between breaks. Combining the test data of action characteristics with the theory of optimal opening distance for current zero crossing, a DC breaking platform for the vacuum circuit breaker was designed, and control strategies were determined, thus completing the 100 kA DC breaking experiment. The vacuum circuit breaker studied in this article meets the technical requirements of the CRAFT quench protection system and provides experience and experimental data for the design of high-current vacuum circuit breakers