Functional proteomics reveals that Slr0237 is a SigE-regulated glycogen debranching enzyme pivotal for glycogen breakdown.

The group 2 σ factor for RNA polymerase SigE plays important role in regulating central carbon metabolism in cyanobacteria. However, the regulation of SigE for these pathways at a proteome level remains unknown. Using a sigE-deficient strain (ΔsigE) of Synechocystis sp. PCC 6803 and quantitative proteomics, we found that SigE depletion induces differential protein expression for sugar catabolic pathways including glycolysis, oxidative pentose phosphate (OPP) pathway, and glycogen catabolism. Two glycogen debranching enzyme homologues Slr1857 and Slr0237 are found differentially expressed in ΔsigE. Glycogen determination indicated that Δslr0237 accumulated glycogen under photomixotrophic condition but was unable to utilize these reserves in the dark, whereas Δslr1857 accumulates and utilizes glycogen in a similar way as the WT strain does in the same condition. These results suggest that Slr0237 plays the major role as the glycogen debranching enzyme in Synechocystis.

Correction: Quantum and semiclassical studies of nonadiabatic electronic transitions between N(4S) and N(2D) by collisions with N2.

Correction for 'Quantum and semiclassical studies of nonadiabatic electronic transitions between N(4S) and N(2D) by collisions with N2' by Dandan Lu et al., Phys. Chem. Chem. Phys., 2023, 25, 15656-15665, https://doi.org/10.1039/D3CP01429K.

Theoretical study of the O(3P) + CN(X2Σ+) → CO(X1Σ+) + N(2D)/N(4S) reactions.

The barrierless exothermic reactions between atomic oxygen and the cyano radical, O(3P) + CN(X2Σ+) → CO(X1Σ+) + N(2D)/N(4S), play a significant role in combustion, astrochemistry, and hypersonic environments. In this work, their dynamics and kinetics are investigated using both wave packet (WP) and quasi-classical trajectory (QCT) methods on recently developed potential energy surfaces of the 12A', 12A,″ and 14A″ states. The product state distributions in the doublet pathway obtained with the WP method for a few partial waves show extensive internal excitation in the CO product. This observation, combined with highly oscillatory reaction probabilities, signals a complex-forming mechanism. The statistical nature of the reaction is confirmed by comparing the WP results with those from phase space theory. The calculated rate coefficients using the WP (with a J-shifting approximation) and QCT methods exhibit agreement with each other near room temperature, 1.77 × 10-10 and 1.31 × 10-10 cm3 molecule-1 s-1, but both are higher than the existing experimental results. The contribution of the quartet pathway is small at room temperature due to a small entrance channel bottleneck. The QCT rate coefficients are further compared with experimental results above 3000 K, and the agreement is excellent.

Methods of privacy-preserving genomic sequencing data alignments.

Genomic data alignment, a fundamental operation in sequencing, can be utilized to map reads into a reference sequence, query on a genomic database and perform genetic tests. However, with the reduction of sequencing cost and the accumulation of genome data, privacy-preserving genomic sequencing data alignment is becoming unprecedentedly important. In this paper, we present a comprehensive review of secure genomic data comparison schemes. We discuss the privacy threats, including adversaries and privacy attacks. The attacks can be categorized into inference, membership, identity tracing and completion attacks and have been applied to obtaining the genomic privacy information. We classify the state-of-the-art genomic privacy-preserving alignment methods into three different scenarios: large-scale reads mapping, encrypted genomic datasets querying and genetic testing to ease privacy threats. A comprehensive analysis of these approaches has been carried out to evaluate the computation and communication complexity as well as the privacy requirements. The survey provides the researchers with the current trends and the insights on the significance and challenges of privacy issues in genomic data alignment.

Unusual Kinetics Induced by Ligands in Rhodium(III)-Catalyzed Dehydrogenative Olefination Reactions.

In this study, we investigate the effects of ligands on C-H activation during rhodium(III)-catalyzed C-H bond olefination reactions using well-defined [CpXRhIII] catalytic systems with three representative CpX (Cp (η5-C5H5), CpCF3 (η5-C5Me4CF3), and Cp* (η5-C5Me5)) ligands. Our results demonstrate that C-H activation as the rate-limiting step is significantly influenced by the steric properties of the CpX ligands. Moreover, we observe a dramatic acceleration of the simple [CpRhIII]-catalyzed C-H olefination reaction with acid coproducts such as HOAc, implying an autocatalytic C-H activation process.

Quantum and semiclassical studies of nonadiabatic electronic transitions between N(4S) and N(2D) by collisions with N2.

The dynamics and kinetics of spin-forbidden transitions between N(2D) and N(4S) via collisions with N2 molecules are investigated using a quantum wave packet (WP) method and the semi-classical coherent switches with decay of mixing (CSDM) method. These electronic transition processes are competing with exchange reaction channels on both the doublet and quartet potential energy surfaces. The WP and CSDM quenching rate coefficients are found in reasonable agreement with each other, and both reproduce the previous theoretical results. For the excitation process, the agreement between the two approaches is dependent on the treatment of the zero-point energy (ZPE) in the product, because the high endoergicity of this process leads to severe violation of the vibrational ZPE. The Gaussian-binning (GB) method is found to improve the agreement with the quantum result. The excitation rate coefficients are found to be two orders of magnitude smaller than that of the adiabatic exchange reaction, underscoring the inefficient intersystem crossing due to the weak spin-orbit coupling between the two spin manifolds of the N3 system.

Quantum and Semiclassical Dynamics of Nonadiabatic Electronic Excitation of C(3P) to C1D) by Hyperthermal Collisions with N2.

The dynamics and kinetics of nonadiabatic excitation of C(3P) to C(1D) induced by hyperthermal collisions with N2 molecules are investigated using a quantum mechanical and two semiclassical nonadiabatic methods. The full-dimensional interaction potential energy surfaces and spin-orbit coupling, which facilitates the spin-forbidden process, are represented by a recently constructed diabatic potential energy matrix. The multistate quantum dynamics for selected partial waves found small transition probabilities due to the weak spin-orbit coupling. The spin-flip transition is the most favored near the threshold due to effective curve crossing. Strong oscillations are also found in the probabilities, which are attributable to resonances supported by the deep well in the singlet-state potential. Vibrational state-specified rate coefficients are reported from J-shifted quantum dynamics calculations, and they follow the Arrhenius form. Vibrational excitation in the N2 collision partner is found to increase the excitation rate at low temperatures, but the trend is reversed at high temperatures. The two semiclassical methods qualitatively reproduce the quantum rate coefficients.

Formation of N(2D) from Hyperthermal Collisions between O(3P) and NO(X2Π).

Hyperthermal collisions between O(3P) and NO(X2Π) could lead to the formation of the first electronically excited atomic nitrogen (N(2D)), which plays a key role in plasma formation in shock-heated air. This process is facilitated mainly by four doublet states, and to a much lesser extent by two quartet states. In this work, we report quasi-classical trajectory studies of this reactive process using the four analytical adiabatic potential energy surfaces for the doublet states developed previously from fitting high-level ab initio data. The reactions were found to be strongly enhanced by vibrational excitation of the NO reactant, consistent with the existence of potential energy barriers in the exit channel. Despite the large endothermicity of the reaction, the rate coefficient is significant at high temperatures, suggesting a possible role of this reaction in the hyperthermal kinetics in the shock layer of a hypersonic (re)entry vehicle.

Hyperthermal Dynamics and Kinetics of the C(3P) + N2(X1Σg+) → CN(X2Σ+) + N(4S) Reaction.

The hyperthermal dynamics and kinetics of the title reaction, which plays an important role in hypersonic chemistry for atmospheric entry vehicles, are investigated using quasi-classical trajectory methods on a recently developed ground electronic state potential energy surface. The dynamics calculations indicated that the reaction follows a complex-forming mechanism, despite its large endoergicity. The calculated differential cross section is forward-backward symmetric, consistent with a long-lived reaction intermediate supported by the NCN potential well. The lifetime of the reaction complex is sufficiently long that the vibrational distribution of the CN product can be predicted by the phase space theory. The calculated vibrational state specific and thermal rate coefficients follow the Arrhenius behavior, and the agreement with existing low-temperature experimental thermal rate coefficients is satisfactory. Extrapolations to high temperatures relevant to hypersonic conditions are provided.

Experimental and Theoretical Studies of Hyperthermal N + O2 Collisions.

The dynamics of hyperthermal N(4S) + O2 collisions were investigated both experimentally and theoretically. Crossed molecular beams experiments were performed at an average center-of-mass (c.m.) collision energy of ⟨Ecoll⟩ = 77.5 kcal mol-1, with velocity- and angle-resolved product detection by a rotatable mass spectrometer detector. Nonreactive (N + O2) and reactive (NO + O) product channels were identified. In the c.m. reference frame, the nonreactively scattered N atoms and reactively scattered NO molecules were both directed into the forward direction with respect to the initial direction of the reagent N atoms. On average, more than 90% of the available energy (⟨Eavl⟩ = 77.5 kcal mol-1) was retained in translation of the nonreactive products (N + O2), whereas a much smaller fraction of the available energy for the reactive pathway (⟨Eavl⟩ = 109.5 kcal mol-1) went into translation of the NO + O products, and the distribution of translational energies for this channel was broad, indicating extensive internal excitation in the nascent NO molecules. The experimentally derived c.m. translational energy and angular distributions of the reactive products suggested at least two dynamical pathways to the formation of NO + O. Quasiclassical trajectory (QCT) calculations were performed with a collision energy of Ecoll = 77 kcal mol-1 using two sets of potential energy surfaces, denoted as PES-I and PES-II, and these theoretical results were compared to each other and to the experimental results. PES-I is a reproducing kernel Hilbert space (RKHS) representation of multireference configurational interaction (MRCI) energies, while PES-II is a many-body permutation invariant polynomial (MB-PIP) fit of complete active space second order perturbation (CASPT2) points. The theoretical investigations were both consistent with the experimental suggestion of two dynamical pathways to produce NO + O, where reactive collisions may proceed on the doublet (12A') and quartet (14A') surfaces. When analyzed with this theoretical insight, the experimental c.m. translational energy and angular distributions were in reasonably good agreement with those predicted by the QCT calculations, although minor differences were observed which are discussed. Theoretical translational energy and angular distributions for the nonreactive N + O2 products matched the experimental translational energy and angular distributions almost quantitatively. Finally, relative yields for the nonreactive and reactive scattering channels were determined from the experiment and from both theoretical methods, and all results are in reasonable agreement.

A Systematic Survey of the Light/Dark-dependent Protein Degradation Events in a Model Cyanobacterium.

Light is essential for photosynthetic organisms and is involved in the regulation of protein synthesis and degradation. The significance of light-regulated protein degradation is exemplified by the well-established light-induced degradation and repair of the photosystem II reaction center D1 protein in higher plants and cyanobacteria. However, systematic studies of light-regulated protein degradation events in photosynthetic organisms are lacking. Thus, we conducted a large-scale survey of protein degradation under light or dark conditions in the model cyanobacterium Synechocystis sp. PCC 6803 (hereafter referred to as Synechocystis) using the isobaric labeling-based quantitative proteomics technique. The results revealed that 79 proteins showed light-regulated degradation, including proteins involved in photosystem II structure or function, quinone binding, and NADH dehydrogenase. Among these, 25 proteins were strongly dependent on light for degradation. Moreover, the light-dependent degradation of several proteins was sensitive to photosynthetic electron transport inhibitors (DCMU and DBMIB), suggesting that they are influenced by the redox state of the plastoquinone (PQ) pool. Together, our study comprehensively cataloged light-regulated protein degradation events, and the results serve as an important resource for future studies aimed at understanding light-regulated processes and protein quality control mechanisms in cyanobacteria.

Putrescine (1,4-Diaminobutane) enhances antifungal activity in postharvest mango fruit against Colletotrichum gloeosporioides through direct fungicidal and induced resistance mechanisms.

Anthracnose decay caused by Colletotrichum gloeosporioides greatly shortens the shelf life and commercial quality of mango fruit. Putrescine (1,4-Diaminobutane) is involved in modulating plant defense to various environmental stresses. In this research, in vivo and in vitro tests were used to explore the antifungal activity and the underlying mechanism of putrescine against C. gloeosporioides in mango fruit after harvested. In vivo tests suggested that putrescine markedly delayed the occurrence of disease and limited the spots expansion on inoculated mango fruit. Further analysis exhibited that putrescine treatment enhanced disease resistance, along with enhanced activities of chitinase (CHI), ß-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), cinnamate-4-hydroxylase (C4H), 4-coumarate coenzyme A ligase (4CL), polyphenol oxidase (PPO) and the accumulation of lignin, flavonoid, phenolics, and anthocyanin in infected mango fruit. In addition, in vitro tests showed that putrescine exerted strongly antifungal activity against C. gloeosporioides. Putrescine induced the production of reactive oxygen species (ROS) and severe lipid peroxidation damage in C. gloeosporioides mycelia, resulting in the leakage of soluble protein, soluble sugar, nucleic acids, K+ and Ca2+ of C. gloeosporioides mycelia. The mycelium treated with putrescine showed severe deformity and shrinkage, and even cracking. Taken together, putrescine could effectively reduce the incidence rate and severity of anthracnose disease possibly through direct fungicidal effect and indirect induced resistance mechanism, thus showing great potential to be applied to disease control.

Reactive and Nonreactive Collisions between NO(X2Π) and O(3P) under Hyperthermal Conditions.

Quasiclassical trajectory calculations are performed for hyperthermal collisions between NO(X2Π) and O(3P) on recently developed potential energy surfaces for the lowest doublet and quartet states of the NO2 system. Three product channels are investigated, and their branching fractions are in reasonably good agreement with the recent crossed molecular beam study at 84 kcal/mol of collision energy. The dominant inelastic channel has a strong forward scattering bias and a high translational energy distribution with limited internal excitation in the scattered NO. The exchange channel has significantly higher NO internal excitation and is also forward biased. The abstraction channel producing internally excited O2 has the smallest branching fraction and a broader angular distribution also with a forward peak. The angular and translational energy distributions in the three channels are consistent with experiment, but the agreement is not always quantitative. The sources of the differences are discussed. Finally, the impact of NO vibrational excitation on the reactive channels and the corresponding rate coefficients are reported.

Distribution of MITE family Monkey King in rapeseed (Brassica napus L) and its influence on gene expression.

Miniature inverted-repeat transposable elements (MITEs) are a group of class II transposable elements. The MITE Monkey King (MK) was first discovered upstream of BnFLC.A10. In this study, genome resequencing of four selected B. napus accessions, revealed more than 4000 distributed copies of MKs constituting ~2.4 Mb of the B. napus genomic sequence and caused 677 polymorphisms among the four accessions. MK -polymorphism-related markers across 128 natural and 58 synthetic accessions revealed more polymorphic MKs in natural than synthetic accessions. Ten MK -induced indels significantly affected the expression levels of the nearest gene based on RNAseq analysis, six of these effects were subsequently confirmed using qRT-PCR. Decreased expression pattern of MK -derived miRNA-bna-miR6031 was also observed under various stress treatments. Further research focused on the MITE families should promote not only our understanding of gene regulatory networks but also inform crop improvement efforts.

Non-Photochemical Quenching: From Light Perception to Photoprotective Gene Expression.

Light is essential for photosynthesis but light levels that exceed an organism's assimilation capacity can cause serious damage or even cell death. Plants and microalgae have developed photoprotective mechanisms collectively referred to as non-photochemical quenching to minimize such potential damage. One such mechanism is energy-dependent quenching (qE), which dissipates excess light energy as heat. Over the last 30 years, much has been learned about the molecular mechanism of qE in green algae and plants. However, the steps between light perception and qE represented a gap in our knowledge until the recent identification of light-signaling pathways that function in these processes in the green alga Chlamydomonas reinhardtii. In this review, we summarize the high light and UV-mediated signaling pathways for qE in Chlamydomonas. We discuss key questions remaining about the pathway from light perception to photoprotective gene expression in Chlamydomonas. We detail possible differences between green algae and plants in light-signaling mechanisms for qE and emphasize the importance of research on light-signaling mechanisms for qE in plants.

Fusion of apoptosis-related protein Cytochrome c with anti-HER-2 single-chain antibody targets the suppression of HER-2+ breast cancer.

Cancer treatment has gradually developed from toxic chemotherapy to targeted therapy with fewer side effects. Approximately 30% of breast cancer patients overexpress human epidermal growth factor receptor 2 (HER-2). Previous studies have successfully produced single-chain antibodies (scFv) targeting HER-2+ breast cancer; however, scFv have poor stability, easy aggregation and a shorter half-life, which have no significant effect on targeting therapy. Moreover, scFv has been considered as a drug delivery platform that can kill target cells by effector molecules. However, the functional killing domains of immunotoxins are mainly derived from plant or bacterial toxins, which have a large molecular weight, low tissue permeability and severe side effects. To address these concerns, we designed several apoptotic immune molecules to replace exogenous toxins using endogenous apoptosis-related protein DNA fragmentation factor 40 (DFF40) and tandem-repeat Cytochrome c base on caspase-3 responsive peptide (DEVD). Our results suggest that DFF40 or Cytc fusion scFv specifically targets HER-2 overexpressing breast cancer cells (SK-BR-3 and BT-474) rather than HER-2 negative cells (MDA-MB-231 and MCF-7). Following cellular internalization, apoptosis-related proteins inhibited tumour activity by initiating endogenous apoptosis pathways, which significantly reduced immunogenicity and toxic side effects. Therefore, we suggest that immunoapoptotic molecules may become potential drugs for targeted immunotherapy of breast cancer.

Vibrational energy pooling via collisions between asymmetric stretching excited CO2: a quasi-classical trajectory study on an accurate full-dimensional potential energy surface.

In low temperature plasmas, energy transfer between asymmetric stretching excited CO2 molecules can be highly efficient, which leads to further excitation (and de-excitation) of the CO2 molecules: CO2(vas) + CO2(vas) → CO2(vas + 1) + CO2(vas - 1). Through such a vibrational ladder climbing mechanism, CO2 can be activated and eventually dissociates. To gain mechanistic insight of such processes, a full-dimensional accurate potential energy surface (PES) for the CO2 + CO2 system is developed using the permutational invariant polynomial-neural network method based on CCSD(T)-F12a/AVTZ energies at about 39 000 geometries. This PES is used in quasi-classical trajectory (QCT) studies of the vibrational energy transfer between CO2 molecules excited in the asymmetric stretching mode. A machine learning algorithm is used to determine state-specific rate coefficients for the vibrational transfer processes from a limited data set. In addition to the CO2(vas + 1) + CO2(vas - 1) channel, the QCT simulations revealed significant contributions from the CO2(vas + 2,3) + CO2(vas - 2,3) channels, particularly at low collision energies/temperatures. These multi-vibrational-quantum processes are attributed to enhanced energy flow in the collisional complex formed by enhanced dipole-dipole interaction between asymmetric stretching excited CO2 molecules.

Phase separation in plants: New insights into cellular compartmentalization.

A fundamental challenge for cells is how to coordinate various biochemical reactions in space and time. To achieve spatiotemporal control, cells have developed organelles that are surrounded by lipid bilayer membranes. Further, membraneless compartmentalization, a process induced by dynamic physical association of biomolecules through phase transition offers another efficient mechanism for intracellular organization. While our understanding of phase separation was predominantly dependent on yeast and animal models, recent findings have provided compelling evidence for emerging roles of phase separation in plants. In this review, we first provide an overview of the current knowledge of phase separation, including its definition, biophysical principles, molecular features and regulatory mechanisms. Then we summarize plant-specific phase separation phenomena and describe their functions in plant biological processes in great detail. Moreover, we propose that phase separation is an evolutionarily conserved and efficient mechanism for cellular compartmentalization which allows for distinct metabolic processes and signaling pathways, and is especially beneficial for the sessile lifestyle of plants to quickly and efficiently respond to the changing environment.

Evaluation of the Potential Risk of Advanced Peak Determination in Distorting Isobaric Labeling-Based Single-Shot Proteome Quantitation.

The recent development and implementation of the advanced peak determination (APD) algorithm with MS instrument dramatically increased the sampling of low abundance features for MS/MS fragmentation. After in-depth evaluation, it is found that with APD on, many chimeric spectra are acquired through co-fragmentation of high abundance contaminants with low abundance targets, and such co-fragmentations are largely avoided when APD is off. To evaluate whether such a co-fragmentation could significantly distort the accuracy of the isobaric-labeling based quantitation of the low abundance target, a single-shot TMT experiment is performed using a two-proteome model, whereby each TMT channel contains premixed peptides from human and a cyanobacterium with a known ratio. Unexpectedly, it is found that APD does not significantly distort TMT ratios, probably because the majority of the APD-specific chimeric spectra are not identifiable. Nevertheless, a few examples of significant distortion of TMT ratios of low abundance peptides caused by APD is found through manual inspection, and suggests that APD should be off in a single-shot TMT experiment to avoid the laborious and time-costing manual inspection.

Baicalein inhibits non-small-cell lung cancer invasion and metastasis by reducing ezrin tension in inflammation microenvironment.

Baicalein, a flavonoid phytochemical, has been shown to be effective as an anti-metastatic agent for various cancers, especially for non-small-cell lung cancer (NSCLC). However, the underlying mechanism of how baicalein targets cellular processes during NSCLC cell invasion and metastasis remains elusive. In this study, we found that non-cytotoxic concentrations of baicalein still retained anti-dissemination activity both in vitro and in vivo. Using a genetic encoding tension probe based on Förster resonance energy transfer (FRET) theory, baicalein was shown to significantly decrease ezrin tension by downregulating cellular ezrin S-nitrosylation (SNO) levels in NSCLC cells in the inflammatory microenvironment. Decreased ezrin tension inhibited the formation of an aggressive phenotype of NSCLC cell and leader cell in collective migration, and subsequently suppressed NSCLC dissemination. Baicalein restrained SNO-mediated ezrin tension by decreasing iNOS expression levels. Overall this study demonstrates the novel mechanism used by baicalein to suppress NSCLC invasion and metastasis from a mechanopharmacology perspective and illustrates a new direction for drug development.