Post-translational Regulation Research Articles

UBIQUITIN-CONJUGATING ENZYME34 mediates pyrophosphatase AVP1 turnover and regulates abiotic stress responses in Arabidopsis.

Understanding the molecular mechanisms of abiotic stress responses in plants is instrumental for the development of climate-resilient crops. Key factors in abiotic stress responses, such as the proton- pumping pyrophosphatase (AVP1), have been identified, but their function and regulation remain elusive. Here, we explored the post-translational regulation of AVP1 by the ubiquitin-conjugating enzyme UBC34 and its relevance in the salt stress and phosphate starvation responses of Arabidopsis (Arabidopsis thaliana). Through in vitro and in vivo assays, we established that UBC34 interacts with and ubiquitylates AVP1. Mutant lines in which UBC34 was downregulated showed higher tolerance to salt and low inorganic phosphate (Pi) stresses, while we observed the opposite for plants overexpressing UBC34. Our results showed that UBC34 co-localizes with AVP1, and AVP1 activity is enhanced in the plasma membrane fractions of ubc34 mutants, indicating that UBC34 mediates the turnover of plasma membrane-localized AVP1. We also observed that UBC34 affects the apoplastic pH, but not the vacuolar pH of root cells. Based on our results, we propose a mechanistic model in which UBC34 mediates AVP1 turnover at the plasma membrane of root epidermal cells. Downregulation of UBC34 under salt and phosphate starvation conditions enhances AVP1 activity, leading to a higher proton gradient available for sodium sequestration and phosphate uptake.

Unravelling malaria latency: parasite intrinsic and environmental factors influencing dormant liver stages.

Hypnozoites - dormant Plasmodium parasites in the liver - can cause relapse infections and form a major obstacle to malaria eradication. The mechanisms controlling dormancy remain poorly understood, but hypnozoite formation and reactivation is likely regulated by a combination of parasite intrinsic factors and external stimuli. We reviewed current knowledge of Plasmodium dormancy and drew parallels with dormancy in other parasites and life-cycle stages. Epigenetic, post-transcriptional, or post-translational regulation probably jointly control hypnozoite dormancy at the intrinsic level. Additionally, environmental factors, such as vector availability, host wellbeing, and tissue microenvironment, could be instrumental to hypnozoite reactivation. A better understanding of how external stimuli influence the intrinsic reactivation switch at a mechanistic level will be required to expand the limited toolset to combat relapsing malaria.

Modification of peroxidase activity and proteome in maize exposed to cadmium in the presence of galactoglucomannan oligosaccharides.

We tested the effects of galactoglucomannan oligosaccharides (GGMOs) and/or cadmium (Cd) on peroxidase activity and the proteome in maize (Zea mays L.) roots and leaves. Our previous work confirmed that GGMOs ameliorate the symptoms of Cd stress in seedlings. Here, the plants were hydroponically cultivated for 7 days, and the protein content and peroxidase activity were estimated in intracellular, neutral cell wall, and acidic cell wall protein fractions. The peroxidase activity varied between the plant organs as well as among the fractions and treatments. The GGMOs in the presence of Cd did not significantly influence content of peroxidases but modulated their activity, which implies posttranslational regulation. The changes in the content of various proteins (e.g., related to the defence reactions, cell wall structure/metabolism, and activation of plant hormones) caused by GGMOs and Cd indicate possible protective mechanisms that improve the vitality of maize seedlings exposed to metal stress. GGMOs partially reverted Cd-induced protein disbalance, which was a reoccurring phenomenon of mitigation in leaves.

Regulation of Histone Acetylation Modification on Biosynthesis of Secondary Metabolites in Fungi.

The histone acetylation modification is a conservative post-translational epigenetic regulation in fungi. It includes acetylation and deacetylation at the lysine residues of histone, which are catalyzed by histone acetyltransferase (HAT) and deacetylase (HDAC), respectively. The histone acetylation modification plays crucial roles in fungal growth and development, environmental stress response, secondary metabolite (SM) biosynthesis, and pathogenicity. One of the most important roles is to regulate the gene expression that is responsible for SM biosynthesis in fungi. This mini-review summarized the regulation of histone acetylation modification by HATs and HDACs on the biosynthesis of SMs in fungi. In most cases, histone acetylation by HATs positively regulated the biosynthesis of fungal SMs, while HDACs had their negative regulations. Some HATs and HDACs were revealed to regulate fungal SM biosynthesis. Hda1 was found to be the most efficient regulator to affect the biosynthesis of SMs in fungi. The regulated fungal species were mainly from the genera of Aspergillus, Calcarisporium, Cladosporium, Fusarium, Monascus, Penicillium, and Pestalotiopsis. With the strategy of histone acetylation modification, the biosynthesis of some harmful SMs will be inhibited, while the production of useful bioactive SMs will be promoted in fungi. The subsequent research should focus on the study of regulatory mechanisms.

From ductal carcinoma in situ to invasive breast cancer: the prognostic value of the extracellular microenvironment.

Ductal carcinoma in situ (DCIS) is a noninvasive breast disease that variably progresses to invasive breast cancer (IBC). Given the unpredictability of this progression, most DCIS patients are aggressively managed similar to IBC patients. Undoubtedly, this treatment paradigm places many DCIS patients at risk of overtreatment and its significant consequences. Historically, prognostic modeling has included the assessment of clinicopathological features and genomic markers. Although these provide valuable insights into tumor biology, they remain insufficient to predict which DCIS patients will progress to IBC. Contemporary work has begun to focus on the microenvironment surrounding the ductal cells for molecular patterns that might predict progression. In this review, extracellular microenvironment alterations occurring with the malignant transformation from DCIS to IBC are detailed. Not only do changes in collagen abundance, organization, and localization mediate the transition to IBC, but also the discrete post-translational regulation of collagen fibers is understood to promote invasion. Other extracellular matrix proteins, such as matrix metalloproteases, decorin, and tenascin C, have been characterized for their role in invasive transformation and further demonstrate the prognostic value of the extracellular matrix. Importantly, these extracellular matrix proteins influence immune cells and fibroblasts toward pro-tumorigenic phenotypes. Thus, the progressive changes in the extracellular microenvironment play a key role in invasion and provide promise for prognostic development.

Building permits-control of type IV pilus assembly by PilB and its cofactors.

Many bacteria produce type IV pili (T4P), surfaced-exposed protein filaments that enable cells to interact with their environment and transition from planktonic to surface-adapted states. T4P are dynamic, undergoing rapid cycles of filament extension and retraction facilitated by a complex protein nanomachine powered by cytoplasmic motor ATPases. Dedicated assembly motors drive the extension of the pilus fiber into the extracellular space, but like any machine, this process is tightly organized. These motors are coordinated by various ligands and binding partners, which control or optimize their functional associations with T4P machinery before cells commit to the crucial first step of building a pilus. This review focuses on the molecular mechanisms that regulate T4P extension motor function. We discuss secondary messenger-dependent transcriptional or post-translational regulation acting both directly on the motor and through protein effectors. We also discuss the recent discoveries of naturally occurring extension inhibitors as well as alternative mechanisms of pilus assembly and motor-dependent signaling pathways. Given that T4P are important virulence factors for many bacterial pathogens, studying these motor regulatory systems will provide new insights into T4P-dependent physiology and efficient strategies to disable them.

NPR1 promotes blue light–induced plant photomorphogenesis by ubiquitinating and degrading PIF4

Light is a major determinant of plant growth and survival. NONEXPRESSER OF PATHOGENESIS-RELATED GENES 1 (NPR1) acts as a receptor for salicylic acid (SA) and serves as the key regulator of SA-mediated immune responses. However, the mechanisms by which plants integrate light and SA signals in response to environmental changes, as well as the role of NPR1 in regulating plant photomorphogenesis, remain poorly understood. This study shows that SA promotes plant photomorphogenesis by regulating PHYTOCHROME INTERACTING FACTOR 4 (PIF4). Specifically, NPR1 promotes photomorphogenesis under blue light by facilitating the degradation of PIF4 through light-induced polyubiquitination. NPR1 acts as a substrate adaptor for the CULLIN3-based E3 ligase, which ubiquitinates PIF4 at Lys129, Lys252, and Lys428, and leading to PIF4 degradation via the 26S proteasome pathway. Genetically, PIF4 is epistatic to NPR1 in the regulation of blue light-induced photomorphogenesis, suggesting it acts downstream of NPR1. Furthermore, cryptochromes mediate the polyubiquitination of PIF4 by NPR1 in response to blue light by promoting the interaction and ubiquitination between NPR1 and PIF4. Transcriptome analysis revealed that under blue light, NPR1 and PIF4 coordinately regulate numerous downstream genes related to light and auxin signaling pathways. Overall, these findings unveil a role for NPR1 in photomorphogenesis, highlighting a mechanism for posttranslational regulation of PIF4 in response to blue light. This mechanism plays a pivotal role in the fine-tuning of plant development, enabling plants to adapt to complex environmental changes.

Chilling- and dark-regulated photoprotection in Miscanthus, an economically important C4 grass

Tolerance of chilling dictates the geographical distribution, establishment, and productivity of C4 crops. Chilling reduces enzyme rate, limiting the sink for the absorbed light energy leading to the need for quick energy dissipation via non-photochemical quenching (NPQ). Here, we characterize NPQ upon chilling in three Miscanthus accessions representing diverse chilling tolerance in C4 grasses. High chilling tolerant accessions accumulate substantial amounts of zeaxanthin during chilling nights in both field and growth chamber settings. Chilling-induced zeaxanthin accumulation in the dark enhances rate of NPQ induction by 66% in the following morning. Based on our data, the emerging ways for the unique regulation of NPQ include post-translational regulation of violaxanthin de-epoxidase (VDE), VDE cofactor accessibility, and absence of transcriptional upregulation of zeaxanthin conversion back to violaxanthin. In the future, more studies will be required to obtain further evidence for these ways contributions to the chilling-dark regulation of NPQ. Engineering dark accumulation of zeaxanthin will help improve crop chilling tolerance and promote sustainable production by allowing early spring planting to maximize the use of early-season soil moisture. Driving the engineered trait by chilling inducible promoter would ensure the minimization of a trade-off between photoprotection and photosynthesis efficiency.

Signal Peptides and Their Fragments in Post-Translation: Novel Insights of Signal Peptides.

Signal peptides (SPs), peptide sequences located at the N-terminus of newly synthesized proteins, are primarily known for their role in targeting proteins to the endoplasmic reticulum (ER). It has traditionally been assumed that cleaved SPs are rapidly degraded and digested near the ER. However, recent evidence has demonstrated that cleaved SP fragments can be detected in extracellular fluids such as blood flow, where they exhibit bioactivity. In addition, SP fragments are delivered to extracellular fluids via extracellular vesicles such as exosomes and microvesicles, which are important mediators of intercellular communication. These findings suggest that SPs and their fragments may have physiological roles beyond their classical function. This review aims to provide a comprehensive overview of these novel roles and offer new insights into the potential functions of SPs and their fragments in post-translational regulation and intercellular communication.

The PRL2 Phosphatase Upregulates miR-21 through Activation of the JAK2/STAT3 Pathway to Downregulate the PTEN Tumor Suppressor.

The Phosphatases of Regenerating Liver (PRLs) are members of the protein tyrosine phosphatase (PTP) superfamily that play pro-oncogenic roles in cell proliferation, migration, and survival. We previously demonstrated that PRLs can post-translationally downregulate PTEN, a tumor suppressor frequently inactivated in human cancers, by dephosphorylating PTEN at Tyr336, which promotes the NEDD4-mediated PTEN ubiquitination and proteasomal degradation. Here we report that PRLs can also reduce PTEN expression by upregulating MicroRNA-21 (miR-21), which is one of the most frequently overexpressed miRNAs in solid tumors. We observe a broad correlation between PRL and miR-21 levels in multiple human cancers. Mechanistically, PRL2, the most abundant and ubiquitously expressed PRL family member, promotes the JAK2/STAT3 pathway-mediated miR-21 expression by directly dephosphorylating JAK2 at Tyr570. Finally, we confirm that the PRL2-mediated miR-21 expression contributes to its oncogenic potential in breast cancer cells. Our study defines a new functional role of PRL2 in PTEN regulation through a miR-21-dependent post-transcriptional mechanism, in addition to our previously reported NEDD4-dependent post-translational PTEN regulation. Together, these studies further establish the PRLs as negative regulators of PTEN.

Deep phosphoproteomics of Klebsiella pneumoniae reveals HipA-mediated tolerance to ciprofloxacin.

Klebsiella pneumoniae belongs to the group of bacterial pathogens causing the majority of antibiotic-resistant nosocomial infections worldwide; however, the molecular mechanisms underlying post-translational regulation of its physiology are poorly understood. Here we perform a comprehensive analysis of Klebsiella phosphoproteome, focusing on HipA, a Ser/Thr kinase involved in antibiotic tolerance in Escherichia coli. We show that overproduced K. pneumoniae HipA (HipAkp) is toxic to both E. coli and K. pneumoniae and its toxicity can be rescued by overproduction of the antitoxin HipBkp. Importantly, HipAkp overproduction leads to increased tolerance against ciprofloxacin, a commonly used antibiotic in the treatment of K. pneumoniae infections. Proteome and phosphoproteome analyses in the absence and presence of ciprofloxacin confirm that HipAkp has Ser/Thr kinase activity, auto-phosphorylates at S150, and shares multiple substrates with HipAec, thereby providing a valuable resource to clarify the molecular basis of tolerance and the role of Ser/Thr phosphorylation in this human pathogen.

The Biology and Clinical Implications of PCSK7.

Discovered in 1996, PCSK7 is the seventh of the nine-membered proprotein convertase subtilisin-kexin (PCSK) family. This article reviews the various aspects of the multifaceted biology of PCSK7 and what makes it an exciting new target for metabolic dysfunction-associated steatotic liver disease (MASLD) affecting ∼30% of the population globally, dyslipidemia, cardiovascular disease (CVD), and likely cancer/metastasis. We will systematically review and discuss all the available epidemiological data, structural, cell biology, and in vivo evidence that eventually led to the discovery of PCSK7 as a novel post-translational regulator of apolipoprotein B. Interestingly, PCSK7 is the only convertase, other than PCSK9, that exhibits non-canonical/non-enzymatic functions, which will be amply described in this review. The data so far suggested that PCSK7 is a potential safe target in MASLD treatment. This was based on human epidemiological data, as well as mouse Pcsk7 knockout and mRNA translation inhibition using hepatocyte-targeted antisense oligonucleotides following a diet-induced MASLD. Additionally, of all the 9 convertases only the gene deletion of Pcsk7 and/or Pcsk9 in mice leads to healthy and fertile animals with no apparent deleterious consequences, suggesting that their pharmacological targeting is likely safe. Accordingly, the synergistic effects of inhibiting both PCSK7 and PCSK9 in a clinical setting may represent a novel therapy for various diseases. We believe that the current surge in oligonucleotide therapy, with many FDA-approved oligonucleotide-based drugs now available in the clinics, and the urgent need for novel MASLD therapeutics present an opportune moment for this timely review article.

Lactylation as a post-translational regulator of cGAS and immunity

In a recent paper at Nature, Li etal.1 reported that alanine-tRNA synthetases AARS1 and AARS2 are lactate sensors and mediate lactylation of cGAS, leading to its inactivation.

Protein arginine methyltransferase 7 modulators in disease therapy: Current progress and emerged opportunity.

Protein arginine methyltransferase 7 (PRMT7) is an essential epigenetic and post-translational regulator in eukaryotic organisms. Dysregulation of PRMT7 is intimately related to multiple types of human diseases, particularly cancer. In addition, PRMT7 exerts multiple effects on cellular processes such as growth, migration, invasion, apoptosis, and drug resistance in various cancers, making it as a promising target for anti-tumor therapeutics. In this review, we initially provide an overview of the structure and biological functions of PRMT7, along with its association with diseases. Subsequently, we summarized the PRMT inhibitors in clinical trials and the co-crystal structural of PRMT7 inhibitors. Moreover, we also focus on recent progress in the design and development of modulators targeting PRMT7, including isoform-selective and non-selective PRMT7 inhibitors, and the dual-target inhibitors based on PRMT7, from the perspectives of rational design, pharmacodynamics, pharmacokinetics, and the clinical status of these modulators. Finally, we also provided the challenges and prospective directions for PRMT7 targeting drug discovery in cancer therapy.

Interplaying Role of Epigenetic, Exercise and Nutrition in Metabolic Diseases

It is important to understand the interplay of epigenetic changes, exercise and nutrition associated with the development of obesity and type-2 diabetes because it offers insights into how environment and lifestyle factors can influence gene expression without altering the genetic code. This knowledge can lead to the development of optimal interventions and preventive strategies that could potentially decrease the rising rates of obesity and type-2 diabetes. The purpose of this review is to examine the interplay of epigenetic, exercise and nutrition on inflammation and metabolic diseases including obesity and type-2 diabetes, and to acquaint the clinicians and researchers with the current advancing and evolving field of exercise and epigenetics. The proposed mechanisms involved pertain to the development of insulin resistance in peripheral tissues and type-2 diabetes including inflammation. The benefits with regular exercise include improvement in metabolic health occurring through adaptative mechanisms in the adipose tissue, skeletal muscle, and liver as well as improvement in insulin resistance. Physical activity is defined by providing physiological characteristics of physical fitness that includes aerobic or cardiorespiratory fitness, and muscular strength and muscular endurance via regular aerobic exercise or resistance training. Exercise training encompasses training modality, exercise frequency and duration. We discuss the interplay of epigenetic mechanisms in inflammation that may contribute to the current worldwide obesity and type-2 diabetes. Specifically, epigenetic induces a change in phenotype without changes in genotype and thus the epigenome can modify the genome outcome through several processes that include DNA methylation, post-translational histone modification and gene regulation mediated by non-coding RNA (ncRNA) mechanisms that have been correlated with various metabolic and inflammatory diseases.

Dual BACH1 regulation by complementary SCF-type E3 ligases.

Broad-complex, tramtrack, and bric-à-brac domain (BTB) and CNC homolog 1 (BACH1) is a key regulator of the cellular oxidative stress response and an oncogene that undergoes tight post-translational control by two distinct F-box ubiquitin ligases, SCFFBXO22 and SCFFBXL17. However, how both ligases recognize BACH1 under oxidative stress is unclear. In our study, we elucidate the mechanism by which FBXO22 recognizes a quaternary degron in a domain-swapped β-sheet of the BACH1 BTB dimer. Cancer-associated mutations and cysteine modifications destabilize the degron and impair FBXO22 binding but simultaneously expose an otherwise shielded degron in the dimer interface, allowing FBXL17 to recognize BACH1 as a monomer. These findings shed light on a ligase switch mechanism that enables post-translational regulation of BACH1 by complementary ligases depending on the stability of its BTB domain. Our results provide mechanistic insights into the oxidative stress response and may spur therapeutic approaches for targeting oxidative stress-related disorders and cancer.

Proteome-wide forced interactions reveal a functional map of cell-cycle phospho-regulation in S. cerevisiae

ABSTRACT Dynamic protein phosphorylation and dephosphorylation play an essential role in cell cycle progression. Kinases and phosphatases are generally highly conserved across eukaryotes, underlining their importance for post-translational regulation of substrate proteins. In recent years, advances in phospho-proteomics have shed light on protein phosphorylation dynamics throughout the cell cycle, and ongoing progress in bioinformatics has significantly improved annotation of specific phosphorylation events to a given kinase. However, the functional impact of individual phosphorylation events on cell cycle progression is often unclear. To address this question, we used the Synthetic Physical Interactions (SPI) method, which enables the systematic recruitment of phospho-regulators to most yeast proteins. Using this method, we identified several putative novel targets involved in chromosome segregation and cytokinesis. The SPI method monitors cell growth and, therefore, serves as a tool to determine the impact of protein phosphorylation on cell cycle progression.

Regulation of OATP1B1 Transport Function by Concurrent Phosphorylation and Lysine-Acetylation-A Novel Post-Translational Regulation Mechanism

Regulation of OATP1B1 Transport Function by Concurrent Phosphorylation and Lysine-Acetylation-A Novel Post-Translational Regulation Mechanism

The ability of sarA to limit protease production plays a key role in the pathogenesis of Staphylococcus aureus osteomyelitis irrespective of the functional status of agr.

We demonstrate that mutation of the staphylococcal accessory regulator A (sarA) in the USA300 strain LAC limits virulence in a murine osteomyelitis model to a greater extent than mutation of the accessory gene regulator (agr) and that it does so irrespective of the functional status of agr. Protease production was decreased in the agr mutant but increased in sarA and sarA/agr mutants to a degree that limited biofilm formation. Extracellular protein A (eSpa) and full-length extracellular nuclease (Nuc1) were absent in the conditioned medium (CM) from sarA and sarA/agr mutants, and their abundance was restored in both mutants by eliminating protease production. Cytotoxicity of CM for osteoblasts and osteoclasts was also reduced in both mutants. Cytotoxicity was restored in a protease-deficient sarA mutant but not in the protease-deficient sarA/agr mutant. Reduced cytotoxicity was correlated with the reduced abundance of full-length α-toxin, LukF, and LukS in sarA and sarA/agr mutants. The abundance of these toxins in their full-length form was increased in the protease-deficient sarA mutant by comparison to LAC, demonstrating that mutation of sarA increases the production of these toxins but increased protease production limits their abundance in full-length and presumably functional forms. Most importantly, eliminating protease production enhanced the virulence of sarA and sarA/agr mutants, but had no impact in the agr mutant. We conclude that a key factor in the attenuation of LAC sarA and sarA/agr mutants in osteomyelitis is the increased production of extracellular proteases and its impact on virulence factors that contribute to biofilm formation and cytotoxicity.IMPORTANCEThe persistent emergence of antibiotic-resistant strains has rekindled interest in anti-virulence strategies to combat S. aureus infections. Numerous reports describe anti-virulence strategies focusing on key regulatory elements that globally influence virulence factor production, the two most commonly targeted being the accessory gene regulator (agr) and the staphylococcal accessory regulator A (sarA). We demonstrate that mutation of sarA limits virulence to a greater extent than mutation of agr and that this can be attributed to increased protease production in both sarA and sarA/agr mutants. This illustrates the critical role of sarA in protease-mediated post-translational regulation in S. aureus. It also suggests that an inhibitor of sarA would be more effective than an inhibitor of agr in overcoming the therapeutic recalcitrance of osteomyelitis and that such an inhibitor would remain effective even in the context of agr mutants known to arise in vivo during the transition from acute to chronic infection.

FUT8 Regulates Cerebellar Neurogenesis and Development Through Maintaining the Level of Neural Cell Adhesion Molecule Cntn2.

Core fucosylation at N-glycans, which is uniquely catalyzed by fucosyltransferase FUT8, plays essential roles in post-translational regulation of protein function. Aberrant core fucosylation leads to neurological disorders in individuals with congenital glycosylation disorders (CDG). However, the underlying mechanisms for these neurological defects remain largely unknown. In this study, we have showed that FUT8 and fucosylation are abundant in cerebellum. Specific deletion of Fut8 in cerebellar granule neuron progenitors (GNPs) results in the impaired proliferation and differentiation of GNPs, as well as the compromised neuronal development, synaptic physiology and motor coordination. Mechanistically, we have showed that Fut8 deficiency reduced Contactin 2 (Cntn2) expression, a member of neural cell adhesion molecules (NCAMs). Furthermore, ectopic Cntn2 can rescue the neuronal defects induced by Fut8 deficiency. Collectively, our study has revealed the important roles of FUT8 and core fucosylation in regulating cerebellar development and function through modulating Cntn2 expression.