Intrinsic Protein Research Articles

TiO2 nanoparticles alleviates the effects of drought stress in tomato seedlings

Nanoparticles have been widely used in recent years, to increase plant tolerance under stress conditions. In this study, TiO2 nanoparticles (NPs) (100 ppm) were applied to tomato plants exposed to drought, and the changes were investigated by physiological, biochemical and proteomic methods. It was determined that TiO2 NPs treatment increased the relative water content and decreased the proline and malondialdehyde content under drought conditions. As a result of proteomic analysis, it was revealed that the expression of a total of 132 proteins changed as a result of the comparison of the treatment groups (drought vs. control, control-100 vs. control, and drought-100 vs. drought). One of the most striking results of the study was the increase of the amounts of photosynthesis-related proteins and plasmamembrane intrinsic protein in both drought and control groups with TiO2 NP-treatmtent. The up-regulation of plasmamembrane intrinsic protein is very important for preserving the water potential under drought conditions. Taken together, it was observed that the water potential of the plant was preserved, lipid peroxidation decreased under drought conditions with the application of TiO2 nanoparticles, and the expression of proteins related to photosynthesis, energy and antioxidant system increased. This study provided clues to the molecular mechanism of the results of many studies available in the literature about nanoparticle treatment under stress condition and showed that TiO2 nanoparticles have a great potential to be used to increase the stress tolerance of tomato plants under drought conditions.

Aquaporin genes in garden pea and their regulation by the nano-antioxidant fullerol in imbibing embryos under osmotic stress

Aquaporins (AQPs) are known as small membrane intrinsic proteins that help to transport water and certain solutes through biological membranes. AQPs gene families are being extensively studied in major crops, but less investigated in pea (<italic>Pisum sativum</italic> L.), which is an economically significant legume crop with a huge complex genome. Here, we present a genome-wide identification, structural characterization, subcellular localization, and expression profiling of the AQPs in pea with a particular interest in their involvement in nano fullerol-conferred osmotic stress alleviation. We identified 39 full-length aquaporin genes from the pea genome, which were classified into five subfamilies. The protein structure of aquaporins appears to have substrate-specific residues which are conserved in plants, allowing for inference of substrate specificity. In particular, the PsNIP2-2-2 was identified with a Gly-Ser-Gly-Arg (GSGR) selective filter that indicates the ability to uptake silicon. Analysis of tissue transcriptomes revealed preferred expressions of certain <italic>PsAQPs</italic> in the underground, aerial and reproductive organs, respectively. Development-regulated expressions of two <italic>PsTIPs</italic> and two <italic>PsPIPs</italic> in seeds were noticed. RNA-Seq of the imbibing embryos treated with mannitol (M) or mannitol plus 100 mg/L fullerol (MF) revealed two <italic>PsTIPs</italic> being similarly regulated by M or MF, three <italic>PsNIPs</italic> being up-regulated only by M without F, and four other genes that were only regulated under MF condition. To our knowledge, this is the first report on transcriptional regulation of AQPs by fullerols, which adds to our knowledge on the plant-carbon nano-substances interactions.

Herbaceous Peony Polyphenols Extend the Vase Life of Cut Flowers

Herbaceous peony is a potential material for cut flowers, but its short vase life seriously affects the development of cut herbaceous peony flowers industry. In this study, herbaceous peony polyphenols were applied to extend the vase life of cut flowers, and the results indicated that 8% mass concentration of herbaceous peony polyphenols increased the superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities; increased the soluble protein content of the cut flowers; and effectively reduced the malondialdehyde (MDA) content. Meanwhile, herbaceous peony polyphenols increased the water balance value of cut flowers. In addition, the observation of microstructures indicated that herbaceous peony polyphenols reduced the blockage mainly caused by Aspergillus spp. at the stem ends and inhibited the growth of Aspergillus spp. Additionally, aquaporin genes (AQPs), including three plasma membrane intrinsic protein genes (PlPIP1;2, PlPIP2;1, and PlPIP2;2) and one intrinsic protein gene (PlNIP), were isolated. PlPIP1;2, PlPIP2;1, and PlPIP2;2, which were induced by polyphenol treatment, had common effects on maintaining the water balance of cut flowers. Therefore, herbaceous peony polyphenols can significantly extend the vase life of cut flowers; these results provide for the application of the theoretical reference of herbaceous peony polyphenols in extending the vase life of cut flowers.

Multi-omics Analyses of Non-GM Tomato Scion Engrafted on GM Rootstocks.

Grafting has been widely applied in agricultural production in order to utilize agriculturally valuable traits. The use of genetically modified (GM) plants for grafting with non-GM crops will soon be implemented to generate chimeric plants (transgrafting)*, and the non-GM edible portions thus obtained could fall outside of the current legal regulations. A number of metabolites and macromolecules are reciprocally exchanged between scion and rootstock, affecting the crop properties as food. Accordingly, the potential risks associated with grafting, particularly those related to transgrafting with GM plants, should be carefully evaluated based on scientific evidence. In this study, we prepared a hetero-transgraft line composed of non-GM tomato scion and GM-tobacco rootstock expressing firefly luciferase. We also prepared a homograft line (both rootstock and scion are from non-GM tomato) and a heterograft line (non-GM tobacco rootstock and non-GM tomato scion). The non-GM tomato fruits were harvested from these grafted lines and subjected to comprehensive characterization by multi-omics analysis. Proteomic analysis detected tobacco-derived proteins from both heterograft and hetero-transgraft lines, suggesting protein transfer from the tobacco rootstock to the tomato fruits. No allergenicity information is available for these two tobacco-derived proteins. The transcript levels of the genes encoding two allergenic tomato intrinsic proteins (Sola l 4.0101 and Sola l 4.0201) decreased in the heterograft and hetero-transgraft lines. Several differences were observed in the metabolic profiles, including α-tomatine and nicotine. The accumulation of tobacco-derived nicotine in the tomato fruits of both heterograft and hetero-transgraft lines indicated that the transfer of unfavorable metabolites from rootstock to scion should be assessed as a food safety concern. Further investigations are needed to clarify whether variable environmental conditions and growth periods may influence the qualities of the non-GM edible parts produced by such transgrafted plants.

Computational and Functional Insights of Protein Misfolding in Neurodegeneration.

Protein folding is the process by which a polypeptide chain self-assembles into the correct three-dimensional structure, so that it ends up in the biologically active, native state. Under conditions of proteotoxic stress, mutations, or cellular aging, proteins can begin to aggregate into non-native structures such as ordered amyloid fibrils and plaques. Many neurodegenerative diseases involve the misfolding and aggregation of specific proteins into abnormal, toxic species. Experimental approaches including crystallography and AFM (atomic force microscopy)-based force spectroscopy are used to exploit the folding and structural characterization of protein molecules. At the same time, computational techniques through molecular dynamics, fold recognition, and structure prediction are widely applied in this direction. Benchmarking analysis for combining and comparing computational methodologies with functional studies can decisively unravel robust interactions between the side groups of the amino acid sequence and monitor alterations in intrinsic protein dynamics with high precision as well as adequately determine potent conformations of the folded patterns formed in the polypeptide structure.

Influence mechanisms of different setting time at low temperature on the gel quality and protein structure of Solenocera crassicornis surimi

The effect of setting time (0–36 h) at 4 °C on the quality changes of shrimp (Solenocera crassicornis) gel was evaluated in this study. The quality of shrimp gel over setting time was evaluated by physical properties including gel strength, drip loss, moisture distribution and microstructure. The physical properties of the shrimp gel were effectively improved after more than 24 h of setting. In addition, the mechanism of the shrimp gel formation was also studied by combining chemical bonds, total sulfhydryl, disulfide bond, intrinsic fluorescence spectra and protein pattern. The results indicated that the main forces affecting the physical properties of the shrimp gel were hydrogen bond and hydrophobic interaction, and the formation of disulfide bond during the setting process was beneficial to the construction of a uniform gel structure. Overall, setting for 24–30 h at 4 °C was an effectual way to improve the shrimp gel qualities.

A novel protein domain is important for photosystem II complex assembly and photoautotrophic growth in angiosperms

Photosystem II (PSII) is a multi-subunit protein complex of the photosynthetic electron transport chain that is vital to photosynthesis. Although the structure, composition, and function of PSII have been extensively studied, its biogenesis mechanism remains less understood. Thylakoid rhodanese-like (TROL) provides an anchor for leaf-type ferredoxin:NADP+ oxidoreductase. Here, we report the chacterizaton of a second type of TROL protein, TROL2, encoded by seed plant genomes whose function has not previously been reported. We show that TROL2 is a PSII assembly cofactor with essential roles in the establishment of photoautotrophy. TROL2 contains a 45-amino-acid domain, termed the chlorotic lethal seedling (CLS) domain, that is both necessary and sufficient for TROL2 function in PSII assembly and photoautotrophic growth. Phylogenetic analyses suggest that TROL2 may have arisen from ancestral TROL1 via gene duplication before the emergence of seed plants and acquired the CLS domain via evolution of the sequence encoding its N-terminal portion. We further reveal that TROL2 (or CLS) forms an assembly cofactor complex with the intrinsic thylakoid membrane protein LOW PSII ACCUMULATION2 and interacts with small PSII subunits to facilitate PSII complex assembly. Collectively, our study not only shows that TROL2 (CLS) is essential for photoautotrophy in angiosperms but also reveals its mechanistic role in PSII complex assembly, shedding light on the molecular and evolutionary mechanisms of photosynthetic complex assemblyin angiosperms.

Dynamical Interaction Analysis of Proteins by a Random Forest-Fragment Molecular Orbital (RF-FMO) Method and Application to Src Tyrosine Kinase

Abstract Identifying key intermolecular (amino acid) interactions is crucial for understanding intrinsic protein functions. In this study, we established an efficient method for discovering key interactions by combining the random forest (RF) method, a machine learning algorithm, and an interaction analysis based on the fragment molecular orbital (FMO) method. We applied this method to Src tyrosine kinase and verified its efficacy. We performed molecular dynamics simulations of both the open and closed forms of Src and selected 50 snapshots for each. Then, pair interaction energy (PIE) or inter-fragment interaction energy (IFIE) analyses were performed using FMO with the van der Waals (vdW)-corrected density functional tight-binding (DFTB) method. Among the 100 × 34453 data sets, we can identify the key amino acid pair regulating the open-close transition. This is consistent with the experimental and theoretical results, indicating the usefulness of the presented method. In contrast to the conventional FMO PIE interaction analysis, in the proposed method, the protein dynamics can be partially included using hundreds of trajectory data.

Mycorrhizal Effects on Growth and Expressions of Stress-Responsive Genes (aquaporins and SOSs) of Tomato under Salt Stress.

Environmentally friendly arbuscular mycorrhizal fungi (AMF) in the soil can alleviate host damage from abiotic stresses, but the underlying mechanisms are unclear. The objective of this study was to analyze the effects of an arbuscular mycorrhizal fungus, Paraglomus occultum, on plant growth, nitrogen balance index, and expressions of salt&amp;nbsp;overly&amp;nbsp;sensitive genes (SOSs), plasma&amp;nbsp;membrane&amp;nbsp;intrinsic&amp;nbsp;protein genes (PIPs), and tonoplast&amp;nbsp;intrinsic&amp;nbsp;protein genes (TIPs) in leaves of tomato (Solanum lycopersicum L. var. Huapiqiu) seedlings grown in 0 and 150 mM NaCl stress. NaCl stress severely inhibited plant growth, but P. occultum inoculation significantly improved plant growth. NaCl stress also suppressed the chlorophyll index, accompanied by an increase in the flavonoid index, whereas inoculation with AMF significantly promoted the chlorophyll index as well as reduced the flavonoid index under NaCl conditions, thus leading to an increase in the nitrogen balance index in inoculated plants. NaCl stress regulated the expression of SlPIP1 and SlPIP2 genes in leaves, and five SlPIPs genes were up-regulated after P. occultum colonization under NaCl stress, along with the down-regulation of only SlPIP1;2. Both NaCl stress and P. occultum inoculation induced diverse expression patterns in SlTIPs, coupled with a greater number of up-regulated TIPs in inoculated versus uninoculated plants under NaCl stress. NaCl stress up-regulated SlSOS2 expressions of mycorrhizal and non-mycorrhizal plants, while P. occultum significantly up-regulated SlSOS1 expressions by 1.13- and 0.45-fold under non-NaCl and NaCl conditions, respectively. It was concluded that P. occultum inoculation enhanced the salt tolerance of the tomato, associated with the nutrient status and stress-responsive gene (aquaporins and SOS1) expressions.

Genome-wide identification of Fagus sylvatica aquaporins and their comparative spring and summer expression profiles

Key messageA total of 45 aquaporins was identified in Fagus sylvatica, 35 of which were differentially expressed in spring and summer in the leaves, phloem and xylem of 11-year-old trees.European beech (Fagus sylvatica) has been widely studied in terms of its water relations and local adaptation. However, to date, the underlying basis conferring adaptation to differences in water availability are unknown. Therefore, we examined the expression of aquaporins in trees of four different beech provenances representing the southern and northern range margins, as well as core populations, grown in a common garden. We sampled their xylem, phloem and leaf tissue, when leaves had fully expanded, and in late summer. A total of 45 aquaporin isoforms were identified in the beech genome, of which 35 were detected across all sampled tissues. In our phylogenetic analysis, beech aquaporins clustered into the five subfamilies found in other woody species. Members of the plasma membrane intrinsic protein subfamily generally displayed the highest levels of expression, followed by tonoplast intrinsic proteins. Isoforms of the remaining subfamilies, Noduline-26-like intrinsic proteins, small basic intrinsic proteins and uncharacterised intrinsic proteins, were expressed at very low to moderate levels. The expression of most isoforms was stable or declined from spring to summer. Leaves followed a different expression profile from that of vascular tissues, whereas both phloem and xylem were found to express the same FsMIPs. Tissue-specific aquaporin expression was very similar amongst the four beech provenances, indicating that there is no inherent difference in the capability of these provenances to regulate aquaporin activity. The general decrease in FsMIP expression toward the end of the growing period indicates that aquaporins are involved in tree water relations and growth.

Colistin Resistance and Management of Drug Resistant Infections.

Colistin resistance is a globalized sensible issue because it has been considered a drug of the last-line resort to treat drug-resistant bacterial infections. The product of the mobilized colistin resistance (mcr) gene and its variants are the significant causes of colistin resistance, which is emerging due to the frequent colistin use in veterinary, and these genes circulate among the bacterial community. Apart from mcr genes, some other intrinsic genes and proteins are also involved in colistin resistance. Researchers focus on the most advanced genomics (whole genome sequencing), proteomics, and bioinformatics approaches to explore the question of colistin resistance. To combat colistin resistance, researchers developed various strategies such as the development of newer drugs, the repurposing of existing drugs, combinatorial treatment by colistin with other drugs, a nano-based approach, photodynamic therapy, a CRISPRi-based strategy, and a phage-based strategy. In this timeline review, we have discussed the development of colistin resistance and its management in developing countries.

Interfacial characteristics of artificial oil body emulsions (O / W) prepared using extrinsic and intrinsic proteins: Inspired by natural oil body

In order to explore the stabilization mechanisms of intrinsic protein (IP) and extrinsic protein (EP) in natural oil-bodies, the effects of adding EP and IP in different relative proportions on the stability, interfacial characteristics, and digestibility of artificial oil body (AOB) emulsions were studied. The stability test results showed that the AOB emulsion prepared using 40% EP/60% IP exhibited the highest ζ-potential (−36.3 mV) and storage stability. According to the results of interface properties, competitive adsorption occurred at the interface in the emulsions prepared using IPs and EPs. Furthermore, by measuring the bioavailability of free fatty acids and curcumin during the digestion process, it was elucidated that 40% EP/60% IP developed a stable interfacial membrane, which facilitated the targeted release of curcumin in the intestine and yielded higher bioavailability. These results lay the foundation for clarifying the stabilization mechanism of natural oil body emulsions from an interfacial perspective.

Pro-efferocytic macrophage membrane biomimetic nanoparticles for the synergistic treatment of atherosclerosis via competition effect

Macrophages participate in many links in the pathological process of atherosclerosis (AS) and the regulation of influence of macrophages at the molecular level might be a new avenue for AS treatment. For this aim, the macrophage membrane biomimetic nanoparticles, derived from macrophage membrane coated SHP1i-loaded liposome NPs (MM@Lips-SHP1i) was designed. Due to the reservation of intrinsic membrane proteins and function from macrophages, the biomimic nanoparticles could effectively evade clearance by the immune system, prolong blood circulation time and actively tend and aggregate to atherosclerotic plaques. More importantly, in the plaque area, MM@Lips-SHP1i nanoparticles could compete with macrophages in vivo to bind with oxidized low-density lipoprotein (oxLDL) and lipopolysaccharide (LPS), reduce uptake of new lipids by macrophages, reduce foam cell formation, and inhibit the expression of pro-inflammatory cytokines. In addition, small molecule inhibitor of SHP-1, the downstream effector molecule of CD47 loaded in macrophage membrane biomimetic nanoparticles could interrupt CD47-SIRPα signal transduction in monocytes and macrophages, thereby enhancing the efferocytosis of macrophages, inhibiting the progression of plaque, achieving synergistic treatment of atherosclerosis. This work focuses on the key process in the formation of AS, macrophage foaming and chronic inflammation, and is based on the fact that macrophage membrane biomimetic nanoparticles can preserve the key surface proteins of macrophages closely related to the formation of AS, providing a new avenue to inhibit the progression of AS by utilizing the biological characteristics of macrophage membrane in macrophage membrane biomimetic nanoparticles.

Aquaporin expression and changes in fatty acid levels in apple leaves subjected to partial root-zone irrigation

ABSTRACT Partial root-zone drying (PRD) can induce changes in levels of the membrane fatty acids, transporter channels such as aquaporins (AQPs) and drought stress-responsive transcription factors (TFs) expression. The apple (Malus domestica (Suckow) Borkh. cv. Braeburn) trees were exposed to different PRD treatments for two consecutive years. The plasma-membrane intrinsic proteins (PIP1;1) and tonoplast-intrinsic proteins (TIP1;1) AQPs presented the highest expression in alternate partial root-zone drying (APRD75) and fixed partial root-zone drying (FPRD75) treatments than FPRD50 and control throughout the entire irrigation season, which was 120-140-fold higher in these treatments during some times. The TFs were highly expressed in APRD75 and FPRD75 treatments. The highest unsaturated fatty acid content was observed in APRD75 and FPRD75 treatments, and the lowest saturated fatty acid content was obtained in FPRD50 in both years. In addition to improving the water use efficiency (WUE) and maintaining the yield, APRD75 and FPRD75 treatments had a higher jasmonic acid (JA) content than the control. These results implied that changes in levels of the membrane fatty acids, AQPs, JA and TFs induced by APRD75 and FPRD75 treatments may be one of the possible ways for apple trees to adapt to the changing environmental conditions caused by PRD.

Biocontrol efficacy of Bacillus velezensis strain YS-AT-DS1 against the root-knot nematode Meloidogyne incognita in tomato plants.

Root-knot nematodes (RKNs; Meloidogyne spp.), one of the most economically important plant-parasitic nematodes (PPNs), cause severe yield and quality losses in agriculture annually. The application of biological control agents is an environmentally safe and effective approach to control RKNs. Here, we report the genomic characteristics of a Bacillus velezensis strain YS-AT-DS1 (Bv-DS1) isolated from the tidal soil, revealing that it has a 4.73 Mb circular chromosome with an average GC-content of 46.43%, 3,977 genes, 86 tRNAs, and 27 rRNAs, and contains secondary metabolite clusters for producing antimicrobial compounds. In vitro assays indicated that Bv-DS1 has not only antagonistic activities against fungal pathogens, but also shows nematicidal activity, with a mortality rate of 71.62% mortality rates in second-stage juvenile (J2s) Meloidogyne incognita. We then focused on the biocontrol efficiency of Bv-DS1 against M. incognita in pot assays. Preinoculation with Bv-DS1 enhanced tomato growth, and significantly reduced the infection rate of J2s, and the number of galls and egg masses on tomato roots. The underlying mechanism in Bv-DS1-induced resistance to M. incognita was further investigated through split-root experiments, and analysing the expression of the genes related to jasmonic acid (JA), salicylic acid (SA), and the tonoplast intrinsic protein (TIP). The results indicated that Bv-DS1 could not activate host systemic-induced resistance (ISR) in the split-root system of tomatoes. Additionally, the expression of JA- (LOX D and MC) and SA- (PAL2 and PR) responsive genes did not change in Bv-DS1-pretreated plants at 3 and 14 days after nematode inoculation. The presented data showed that JA-and SA-dependent pathways were not required for the biocontrol action of the Bv-DS1 against RKN. The TIP genes, responsible for transport of water and small substrates in plants, have previously been shown to negatively regulate the parasitism of PPNs. Surprisingly, Bv-DS1 compromised the downregulation of TIP1.1 and TIP1.3 by M. incognita. Together, our data suggest that Bv-DS1 exhibits a dual effect on plant growth promotion and protection against RKN, possibly related to the regulation of water and solute transport via TIPs. Thus, the Bv-DS1 strain could be used as a biocontrol agent for RKN control in sustainable agriculture.

The Integration of Proteome-Wide PTM Data with Protein Structural and Sequence Features Identifies Phosphorylations that Mediate 14-3-3 Interactions

14-3-3s are abundant proteins that regulate essentially all aspects of cell biology, including cell cycle, motility, metabolism, and cell death. 14-3-3s work by docking to phosphorylated Ser/Thr residues on a large network of client proteins and modulating client protein function in a variety of ways. In recent years, aided by improvements in proteomics, the discovery of 14-3-3 client proteins has far outpaced our ability to understand the biological impact of individual 14-3-3 interactions. The rate-limiting step in this process is often the identification of the individual phospho-serines/threonines that mediate 14-3-3 binding, which are difficult to distinguish from other phospho-sites by sequence alone. Furthermore, trial-and-error molecular approaches to identify these phosphorylations are costly and can take months or years to identify even a single 14-3-3 docking site phosphorylation. To help overcome this challenge, we used machine learning to analyze predictive features of 14-3-3 binding sites. We found that accounting for intrinsic protein disorder and the unbiased mass spectrometry identification rate of a given phosphorylation significantly improves the identification of 14-3-3 docking site phosphorylations across the proteome. We incorporated these features, coupled with consensus sequence prediction, into a publicly available web app, called “14-3-3 site-finder”. We demonstrate the strength of this approach through its ability to identify 14-3-3 binding sites that do not conform to the loose consensus sequence of 14-3-3 docking phosphorylations, which we validate with 14-3-3 client proteins, including TNK1, CHEK1, MAPK7, and others. In addition, by using this approach, we identify a phosphorylation on A-kinase anchor protein-13 (AKAP13) at Ser2467 that dominantly controls its interaction with 14-3-3.

Ice plant root plasma membrane aquaporins are regulated by clathrin-coated vesicles in response to salt stress.

The regulation of root Plasma membrane (PM) Intrinsic Protein (PIP)-type aquaporins (AQPs) is potentially important for salinity tolerance. However, the molecular and cellular details underlying this process in halophytes remain unclear. Using free-flow electrophoresis and label-free proteomics, we report that the increased abundance of PIPs at the PM of the halophyte ice plant (Mesembryanthemum crystallinum L.) roots under salinity conditions is regulated by clathrin-coated vesicles (CCV). To understand this regulation, we analyzed several components of the M. crystallinum CCV complexes: clathrin light chain (McCLC) and subunits μ1 and μ2 of the adaptor protein (AP) complex (McAP1μ and McAP2μ). Co-localization analyses revealed the association between McPIP1;4 and McAP2μ and between McPIP2;1 and McAP1μ, observations corroborated by mbSUS assays, suggesting that AQP abundance at the PM is under the control of CCV. The ability of McPIP1;4 and McPIP2;1 to form homo- and hetero-oligomers was tested and confirmed, as well as their activity as water channels. Also, we found increased phosphorylation of McPIP2;1 only at the PM in response to salt stress. Our results indicate root PIPs from halophytes might be regulated through CCV trafficking and phosphorylation, impacting their localization, transport activity, and abundance under salinity conditions.

Saussurea involucrata PIP2;4 improves growth and drought tolerance in Nicotiana tabacum by increasing stomatal density and sensitivity

Aquaporins, the major facilitators of water transport across membranes, are involved in growth and development and adaptation to drought stress in plants. In this study, a plasma membrane intrinsic protein (SiPIP2;4) was cloned from Saussurea involucrata, a cold-tolerant hardy herb. The expression of SiPIP2;4 increased the stomatal density and sensitivity of tobacco (Nicotiana tabacum), thus, affecting the plant’s growth and resistance to the diverse water environment. The higher stomatal density under well-watered conditions effectively promoted the photosynthetic rate, which led to the rapid growth of transgenic lines. The stomata in the transgenic lines responded more sensitively to the vapor pressure deficit than the wild-type under different levels of ambient humidity. Their stomatal apertures positively correlated with the ambient humidity. Under drought conditions, the overexpression of SiPIP2;4 promoted rapid stomatal closure, reduced water dissipation, and enhanced drought tolerance. These results indicate that SiPIP2;4 regulates the density and sensitivity of plant stomata, thus, playing an important role in balancing plant growth and stress tolerance. This suggests that SiPIP2;4 has the potential to serve as a genetic resource for crop improvement.

Anterior Umbilication of Lens in a Family with Congenital Cataracts Associated with a Missense Mutation of MIP Gene.

Congenital cataracts (CCs) have significant genotypic and phenotypic heterogeneity. The major intrinsic protein (MIP) gene, one of the causative genes of CCs, plays a vital role in maintaining the homeostasis and transparency of the lens. In this study, we identified a unique phenotype of anterior umbilication of the lens in a four-generation pedigree with CCs. All patients in the observed family had nystagmus, nuclear cataracts, and elongated axial lengths compared with their healthy counterparts except for patient I:2, whose axial length was unavailable, and patientII:4, who had total cataracts. We confirmed, using Sanger sequencing based on whole-exon sequencing (WES) data, that all patients carried a heterozygous variant NM_012064.4:c.97C > T (NP_036196.1:p.R33C) in their MIP gene. To our knowledge, 29 variants of the human MIP gene and the relative phenotypes associated with CCs have been identified. Nevertheless, this is the first report on the anterior umbilication of the lens with nuclear or total opacity caused by the c.97C > T (p.R33C) variant in the MIP gene. These results also provide evidence that the elongated axial length might be associated with this variant. This study further confirms the phenotypic heterogeneity of CCs.

Intrinsic protein disorder and conditional folding in AlphaFoldDB.

Intrinsically disordered regions (IDRs) defying the traditional protein structure–function paradigm have been difficult to analyze. The availability of accurate structure predictions on a large scale in AlphaFoldDB offers a fresh perspective on IDR prediction. Here, we establish three baselines for IDR prediction from AlphaFoldDB models based on the recent CAID dataset. Surprisingly, AlphaFoldDB is highly competitive for predicting both IDRs and conditionally folded binding regions, demonstrating the plasticity of the disorder to structure continuum.