Search Results (217)

Mycoplasma bovis is an important pathogen that is associated with respiratory diseases, mastitis, and arthritis in cattle, leading to significant economic losses in the global cattle industry. Most notably in this study, we pioneer the discovery that its secreted effector ENO1 (α-enolase) directly targets host cytoskeletal proteins for metabolic–immune regulation. Using an innovative GST pull-down/mass spectrometry approach, we made the seminal discovery of β-actin (ACTB) as the primary host target of ENO1—the first reported bacterial effector–cytoskeleton interaction mediating metabolic reprogramming. ENO1–ACTB binding depends on a hydrogen bond network involving ACTB’s 117Glu and 372Arg residues. This interaction triggers (1) glycolytic activation via Glut1 upregulation, establishing Warburg effect characteristics (lactic acid accumulation/ATP inhibition), and (2) ROS-mediated activation of dual inflammatory axes (HIF-1α/IL-1β and IL-6/TNF-α). This work establishes three groundbreaking concepts: (1) the first evidence of a pathogen effector hijacking host ACTB for metabolic manipulation, (2) a novel ‘glycolysis–ACTB–ROS-inflammation’ axis, and (3) the first demonstration of bacterial proteins coordinating a Warburg effect with cytokine storms. These findings provide new targets for anti-infection therapies against Mycoplasma bovis. Full article

Iridoids are compounds recognized for their neuroprotective properties and their potential application in the treatment of neurodegenerative diseases. Geniposide (GP) and asperuloside (ASP) are iridoids that have demonstrated some biological activities. In this study, the potential neuroprotective effects of these iridoids were evaluated through in silico and in vivo assays, using Caenorhabditis elegans (C. elegans) strains CF1553 (sod-3::GFP), GA800 (cat::GFP), and CL2166 (gst-4::GFP). The results suggested that neither compound appears to have good passive permeability through the blood–brain barrier (BBB). However, an active transport mechanism involving the glucose transporter GLUT-1 may be present, as both compounds contain glucose in their molecular structure. In addition, they can inhibit the activity of both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). GP at 1 and 2 mM reversed the H₂O₂-induced increase in sod-3 expression, while ASP at 1 and 2 mM reversed the increase in gst-4 expression. Worm survival was more adversely affected by higher concentrations of GP than ASP, although both similarly reduced acetylcholinesterase activity. These findings suggest that GP and ASP exhibit very low toxicity both in silico and in vivo in C. elegans, and positively modulate key enzymes involved in antioxidant pathways, highlighting their potential for neuroprotective applications. Full article

Microplastics (MPs) are a major contaminant in aquatic environments. Due to their size, they are likely to cause deleterious effects. In this study, we assessed the effects of MPs obtained from two commercially available plastics (PP and PET) in the polychaeta Hediste diversicolor after different periods (4 and 28 days). Toxic effects were assessed by measuring burrowing and spontaneous activities, phase I (CYP1A1, 1A2, and 3A4) activities), conjugation metabolism (GSTs), and antioxidant defense (CAT). Behavioral traits and phase I activities were nonresponsive to the presence of both plastics and for the two durations of exposure, indicating that these organisms are not affected by exposure to MPs and do not metabolize them. Conjugation metabolism was inhibited, which may be explained by the MPs’ capability of inhibiting certain enzymes. CAT activity was increased in animals acutely exposed to PP and decreased in animals chronically exposed to PET. This study shows that PP- and PET-MPs do not cause adverse effects on H. diversicolor. Full article

Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive liver disease linked to fibrosis and hepatocellular carcinoma (HCC). Gut-derived lipopolysaccharide (LPS) promotes hepatic inflammation, fibrosis, and angiogenesis through toll-like receptor 4 (TLR4) signaling. This study examined the effects of rifaximin, a non-absorbable, gut-targeted antibiotic, on MASH-related liver fibrosis and early hepatocarcinogenesis, with a focus on the LPS–epiregulin–IL-8–angiogenesis axis.MASH was induced in Fischer 344 rats using a choline-deficient, L-amino acid-defined high-fat diet (CDAHFD). Rifaximin (30 mg/kg/day) was orally administered for 12 weeks. Liver histology, gene expression, intestinal permeability, LPS levels, and angiogenic markers were evaluated. Rifaximin reduced hepatic inflammation, fibrosis, hydroxyproline content, and fibrogenic gene expression. The number and size of GST-P-positive preneoplastic lesions and proliferation-related genes were decreased. Portal LPS levels and Kupffer cell activation declined, with downregulation of Lbp, Cd14, Tlr4, and inflammatory cytokines. Rifaximin decreased hepatic epiregulin and IL-8 expression, attenuated CD34-positive neovascularization, and suppressed proangiogenic gene expression, accompanied by improved intestinal barrier function and reduced gut permeability. Rifaximin mitigates MASH progression by restoring gut barrier integrity, limiting LPS translocation, and inhibiting fibrogenic and angiogenic pathways. These results suggest its potential as a chemopreventive agent in MASH-related hepatocarcinogenesis. Full article

This study investigated the ecotoxicological impacts of environmentally relevant concentrations (0.05, 0.50, and 5.00 mg/L) of nickel nanoparticles (Ni-NPs) by assessing oxidative stress biomarkers. The worm Hediste diversicolor, the bivalve Mytilus spp., and the fish Sparus aurata were chronically exposed to Ni-NPs for 28 days, and glutathione S-transferases (GST), catalase (CAT), and thiobarbituric acid reactive substances (TBARS) levels were measured to evaluate biochemical responses. GST activity increased in H. diversicolor and the liver of S. aurata, suggesting a key role for this enzyme in Ni-NPs detoxification. CAT activity was inhibited in the digestive gland of Mytilus spp. at the highest Ni-NPs concentration, indicating possible disruption of antioxidant defense. TBARS levels rose significantly in the gills of Mytilus spp. exposed to high Ni-NP concentrations, suggesting oxidative damage beyond detoxification capacity. In contrast, TBARS decreased in the digestive gland of Mytilus and in H. diversicolor, possibly due to compensatory upstream antioxidant responses. These findings indicate that each species exhibits distinct adaptive responses to Ni-NP exposure. Overall, this study highlights the need to consider species- and tissue-specific responses when performing ecotoxicological risk assessments of nanomaterials. Full article

The knowledge about the potential toxic effects of microplastics (MPs) combined with herbicides at lower trophic levels is still largely unknown. The present study aimed to evaluate the potential toxic effects of polyethylene terephthalate (PET) and polyamide (PA), isolated or combined with the pesticide glyphosate (GLY), on the microalgae Arthrospira platensis. For this, microalgae were exposed to control, GLY (3 μg/L), PET (0.5 and 1 mg/L), PA (0.5 and 1 mg/L), and the respective mixtures of each MP with GLY, for 12 days. The photosynthetic pigment content, phytochemicals, antioxidants, and enzymatic activity were determined. Cell growth was significantly enhanced on day 4 in the GLY+PA1 group (~80%), compared to the control. At day 12, biomass was significantly higher in the GLY (~25%) and GLY+PET0.5 (~26%) groups relative to the control. Significant effects on the enzymatic and detoxification mechanisms were observed, including increased SOD (PET0.5, p = 0.011) and CarE (GLY, PA and GLY+PA, p < 0.01), and decreased GST in combined exposures, which support stress-induced enzymatic activation and adaptive biochemical responses. Significant effects on phytochemicals and antioxidant activity were also observed, with PET0.5 significantly reducing total carotenoids (~65%), and flavonoids (p < 0.001) and ortho-diphenols (p < 0.05) being decreased in all exposure groups, in comparison to the control group. The decrease in flavonoids and ortho-diphenols, important antioxidant molecules, suggests the depletion of these key compounds under stress. DPPH scavenging activity, a measure of antioxidant potential, was inhibited in the GLY+PA groups, indicating compromised antioxidant defense. Results confirmed that combined stressors elicit distinct and sometimes deleterious responses not predicted by single exposures. Our findings highlight that the combined exposure to glyphosate and MPs significantly disrupts antioxidant defenses and enzymatic activity in A. platensis, indicating potential risks to primary producers in aquatic ecosystems and underscoring the ecological implications of co-contaminant stressors. In fact, the results indicate that MPs can modify herbicide toxicity, posing enhanced risks to microalgal physiology and potentially affecting primary productivity and nutrient cycling in aquatic ecosystems. In turn, negative effects of MPs on microalgae can have serious consequences for food webs, food security, and ecological health. Full article

The widespread use and pseudo-persistent occurrence of the antidepressant citalopram (CIT) could pose a potential ecological risk in the aquatic environment. The message about the bioconcentration and sensitive biomarker identification of CIT at the environmentally relevant concentrations is limited. In this study, an integral evaluation of the phenotypic and biochemical effects of CIT on Daphnia magna (D. magna) was conducted at 0.5 and 10 µg/L. The biomarker screening includes energy metabolism, phototactic behavior, feeding dysfunction, and antioxidant stress responses. The carbohydrate, lipid, and protein content was determined using the assay of anthrone with glucose as standard, thiophosphorate-Vaniline with cholesterol as standard, and Coomassie brilliant blue with serum albumin as standard, respectively. The results showed the bioconcentration equilibrium of CIT reached at the exposure duration of 48 h during the uptake process. At the exposure concentrations of 0.5 and 10 µg/L, the bioconcentration factor of CIT was 571.2 and 67.4 L/kg, respectively. Both protein and lipid content significantly increased at 0.5 µg/L with a 1.78-fold elevation in total energy. Comparatively, the lipid content showed a significant increase at 10 µg/L, while the available total energy rose by 1.25-fold relative to the control group. The phototactic behavior of D. magna exposed to 0.5 µg/L CIT was markedly reduced at 48 h relative to control. In contrast, a significant decrease in phototaxis was observed after 6 h and then a significant increase at 12 h with a continuously obvious decline at 10 µg/L. The filtration rates were increased by 32% compared to controls at 0.5 µg/L, while the stimulatory effects disappeared at 10 µg/L. With regarding to the antioxidant enzyme activities, CIT exposure significantly inhibited the catalase activity both at 0.5 and 10 µg/L, while the glutathione S-transferase activity was obviously induced at 0.5 µg/L and inhibited at 10 µg/L. The expression level of 18s gene was significantly decreased at 10 µg/L. Only the gst gene expression level was significantly increased at 0.5 µg/L, while the 18s and cat gene expression level was obviously inhibited and induced at 10 µg/L. Comprehensively, the responses of the phenotypic traits and energy metabolism of D. magna at various environmental concentrations were sensitive for CIT. This study provided basic data for the risk estimation of CIT in the real freshwater environment. Full article

Haemonchosis caused by the parasitic worm Haemonchus contortus is a major threat to cattle and other ruminants and imposes significant economic losses in the livestock industry. Different medications have been reported; however, these are not reliable now due to mass drug resistance. The current study investigates potential inhibitors of two H. contortus proteins: glutathione S-transferase (GST) and beta-tubulin isotype 1. GST helps the parasite to detoxify harmful substances, while beta-tubulin is essential for the cell division and structure. By using computational approaches, natural compounds were identified to inhibit the selected proteins. The 3D structures of GST and β-tubulin isotype 1 were prepared, and pharmacophore models were generated to search the Molport natural compound library. The lowest binding energy ranged from −6.7 to −10.4 Kcal/mol. Post-docking interactional analyses revealed that Glu45, Arg46, Cys126, Gln131, Lys252, Asn247, and Arg251 residues were the most common interacting residues in β-tubulin isotype 1. Similarly, in GST, Leu99, Asn100, Arg103, Lys107, Glu162, and Met163 were the most common interacting residues. In conclusion, extensive computational analyses including virtual screening, docking, and MD simulations revealed that the compound Molport-039-195-358 might have the ability to control haemonchosis by targeting GST and β-tubulin isotype 1. The in silico studies identified potent compounds by targeting GST and β-tubulin isotype 1 against Haemonchus contortus. The reported findings provide a foundation for the development of novel anthelmintic therapies. Full article

(1) Background: The larvae of Dioryctria sylvestrella typically bore into the shoots and cones of Pinus koraiensis, increasing tree breakage risk and reducing cone yield. (2) Methods: Five Beauveria bassiana strains were evaluated for virulence against fourth-instar larvae. And the levels of T-AOC and MDA in the larvae infected by each strain were measured. To assess larval responses to different strains, we measured the activities of six enzymes (SOD, CAT, POD, PPO, CarE, GST) and the levels of GSH and H₂O₂ in larvae treated with each strain. Additionally, the infection process of highly pathogenic B. bassiana in larvae was explored using scanning electron microscopy (SEM). (3) Results: Strain CGMCC3.2055 demonstrated the highest toxicity to larvae, achieving a cumulative corrected mortality of 80.56% on the 4th day and an LT₅₀ of 3.248 days. The T-AOC of larvae treated with strain CGMCC3.2055 was inhibited within 48 h. The relative MDA content in this group was significantly higher than that in other strain-treated groups at 6, 12, and 24 h. In Bb01-treated larvae, H₂O₂ accumulation at 6 and 24 h post-infection was influenced by POD activity rather than GSH levels; in BbZ1-treated larvae, the activities of CAT and POD were upregulated at 6 and 36 h, while the activity of SOD was downregulated, but the content of H₂O₂ increased significantly, resulting in accumulation; in CFCC81428-treated larvae, a decline in T-AOC coincided with substantial H₂O₂ accumulation over 48 h, while a concomitant increase in GSH content bolstered tolerance to lethal oxidative damage; in CGMCC3.2055-treated larvae, H₂O₂ only accumulated significantly at 24 and 48 h, yet upregulated CAT and POD were insufficient to effectively scavenge the excess H₂O₂; and in bio-21738-treated larvae, SOD-driven dismutation generated substantial H₂O₂ from 12 to 48 h, leading to pronounced accumulation from 6 to 48 h, yet limited upregulation of POD (only at 6 and 12 h) and CAT (only at 12 and 48 h) were insufficient to mitigate H₂O₂ buildup. PPO activity was upregulated within 48 h in all treatment groups except for BbZ1, where no upregulation was observed at 12 and 48 h. GST activity was upregulated in all treatment groups except for CGMCC3.2055, where a downregulation was observed at 12 h post-infection. CarE activity was significantly upregulated within 48 h in both CFCC81428 and CGMCC3.2055 groups; in the Bb01 group, CarE was upregulated only at 6 and 48 h; in the BbZ1 group, CarE was downregulated only at 48 h; and in the bio-21738 group, CarE showed no upregulation at 24 and 48 h. Through SEM, the infection process of the strain CGMCC3.2055 on the surface of the larvae was further determined, which mainly included adhesion, the appearance of bud-like protrusions, the growth of germ tubes along the epidermis and penetration of the epidermis, as well as the colonization of the strain and its emergence from the surface of the larvae. (4) Conclusions: This study first screened the highly pathogenic B. bassiana strain CGMCC3.2055 by evaluating its virulence to larvae and post-infection T-AOC and MDA levels. It also clarified the strain’s infection process and the larvae’s immune responses to various strains. Full article

The bird cherry-oat aphid Rhopalosiphum padi (L.) poses a significant threat to wheat production, resulting in substantial yield reductions. Abamectin and acetamiprid are frequently utilized for management. This study assessed the sublethal effects of abamectin and acetamiprid on R. padi through life table analysis and enzyme activity assays. At 24 h, the LC₁₀ and LC₃₀ values for abamectin to R. padi were 0.063 mg/L and 0.252 mg/L, respectively, while, for acetamiprid, the corresponding values were 0.065 and 0.293 mg/L. The results indicated that exposure to sublethal concentrations of abamectin (AB-LC₁₀) extended the longevity of R. padi F₀ generation, while acetamiprid (AC-LC₁₀ and AC-LC₃₀) decreased it. Furthermore, the fecundity of the F₀ generation was significantly reduced following exposure to AB-LC₃₀, AC-LC₁₀ and AC-LC₃₀. In the F₁ generation, exposure to sublethal concentrations of acetamiprid negatively impacted on R. padi, as evidenced by a significant reduction in longevity; fecundity and population parameters (R₀, r, λ, s_xj, l_x, l_xm_x, v_xj and e_xj). Conversely, sublethal concentrations of abamectin did not significantly affect these parameters. Additionally, population projections revealed a significantly smaller total population size of R. padi in the acetamiprid-exposed group compared to both the abamectin-exposed and control groups. Except these population-level effects, the activities of detoxification enzymes, including cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST) and carboxylesterases (CarE), changed differently after treatments. These results suggest that sublethal concentrations of acetamiprid, but not abamectin, significantly inhibit the population growth of R. padi. These insights are crucial for R. padi control and facilitate the development of effective control strategies that take into account these sublethal effects in integrated pest management strategies targeting R. padi. Full article

Sitophilus zeamais, a major pest of stored grains, causes significant post-harvest losses and challenges effective control. While synthetic insecticides pose risks of resistance and toxicity, essential oils (EOs) offer a safer alternative. However, the insecticidal potential of their individual volatile constituents (VCs) remains largely unexplored. This study evaluated the insecticidal activity of 51 EO-derived volatile compounds (VCs) against S. zeamais, identifying the most toxic ones, optimizing 15 synergistic mixtures, and assessing their effects on key insect enzymes. A structure–activity relationship (SAR) analysis determined functional groups associated with insecticidal activity, while a cluster analysis pre-selected 29 ternary mixtures, later refined using response surface methodology (RSM). Additionally, enzymatic assays explored their impact on detoxification and nervous system enzymes, providing insights into potential mechanisms of action. Among the 51 VCs tested, 37 exhibited significant toxicity, with 11 acting as fumigants and 13 displaying contact toxicity. Monocyclic monoterpenoids with ketone or alcohol functional groups and exocyclic unsaturation demonstrated the highest insecticidal activity via both exposure routes. Notably, pulegone enantiomers were particularly effective (LC₅₀ < 0.1 mg/L, LD₅₀ < 7.5 µg/adult). Among the optimized mixtures, 10 displayed strong insecticidal effects, 8 were active through both routes, and 5 exhibited synergistic fumigant interactions. The most effective formulations were M2 (R-pulegone + S-pulegone + S-carvone, LC₅₀ 0.48 mg/L) and M20 (isopulegone + δ-3-carene, LC₅₀ 2.06 mg/L), showing the strongest fumigant and synergistic effects, respectively. Enzymatic assays revealed that while some compounds mildly inhibited GST and CAT, others, such as δ-3-carene (IC₅₀ 0.19 mg/L), significantly inhibited AChE. Five mixtures exhibited synergistic neurotoxicity, with M20 (IC₅₀ 0.61 mg/L) and M12 (IC₅₀ 0.81 mg/L) emerging as the most potent AChE inhibitors. These findings highlight the potential of plant-derived volatile compounds as bioinsecticides, leveraging synergistic interactions to enhance efficacy, disrupt enzymatic pathways, and mitigate resistance. Full article

This study evaluates the effects of caffeine (CAF) on the bacteria Aliivibrio fischeri, the microalga Raphidocelis subcapitata, the macrophyte Lemna minor, and the larvae of Chironomus riparius, aiming to understand its environmental impact and contribution to ecological risk assessment. Bioluminescence inhibition in A. fischeri (EC₅₀ = 998.5 mg/L) and growth inhibition in R. subcapitata and L. minor (EC₅₀ = 60.1 mg/L and EC₅₀ = 649.2 mg/L, respectively) were observed. For L. minor, reduced catalase (CAT) activity and non-linear responses in glutathione S-transferases (GSTs) were recorded. No significant changes were observed in proline, malondialdehyde (MDA), and pigment contents. In C. riparius, acute mortality (LC₅₀ = 644.5 mg/L) was observed, and growth was significantly affected after 10 days of CAF exposure (EC₅₀ = 81.62 mg/L for fresh biomass). After 10 days of exposure, there was an increase in CAT activity and thiobarbituric acid reactive substances, with TBARS levels both at concentrations ≥82.64 mg/L, and a decrease in GSTs (92.18 mg/L) and acetylcholinesterase (AChE) (≤62.09 mg/L) activities of C. riparius. The results show that CAF exposure affects organisms’ metabolic and physiological functions, with varying sensitivities among species, potentially leading to ecological disturbances in aquatic ecosystems. The hazardous concentration for 5% of species was 4.42 mg/L. Long-term studies are necessary to understand the risk of caffeine under more realistic scenarios. Full article

This research explored the role of aflatoxin oxidase CotA in mitigating aflatoxin B₁ (AFB₁)-induced hepatotoxicity in Japanese quails. A total of 225 female Japanese quails, aged two weeks, were randomly assigned to three dietary groups: a control diet, an AFB₁-contaminated diet, and an AFB₁-contaminated diet supplemented with aflatoxin oxidase CotA for three weeks. The results indicate that quails receiving the AFB₁-contaminated diet exhibited reduced body weight gain, pronounced vacuolar degeneration within hepatocytes, and inflammatory cell infiltration. Additionally, the AFB₁ group demonstrated an increased liver index and elevated serum liver enzyme activities (ALT, AST, and ALP). Supplementation with CotA improved body weight gain and conferred protection against AFB₁-induced liver injury. Furthermore, the addition of CotA significantly enhanced liver antioxidant enzyme activities (T-AOC, GST, GSH-Px, POD, and CAT), reduced hepatic H₂O₂ and MDA levels, and upregulated the mRNA expression levels of genes in the Nrf2 pathway in quails exposed to AFB₁. AFB₁ exposure led to lipid droplet accumulation in liver tissues and elevated serum TG and LDL-C levels. However, the introduction of CotA mitigated AFB₁-induced alterations in lipid metabolism. Furthermore, dietary supplementation with CotA inhibited AFB₁-induced hepatocyte apoptosis and decreased the mRNA expression of apoptosis-related genes, including Bax, caspase-9, and caspase-3. Notably, the AFB₁+CotA group exhibited a significant reduction in AFB₁ residues and AFB₁-DNA adducts in quail liver tissues compared to the AFB₁ group. These findings indicate that aflatoxin oxidase CotA holds promise as a feed additive to alleviate AFB₁-induced hepatotoxicity. Full article

Myosin phosphatase (MP) holoenzyme consists of protein phosphatase-1 (PP1) catalytic subunit (PP1c) associated with myosin phosphatase target subunit-1 (MYPT1) and it plays an important role in mediating the phosphorylation of the 20 kDa light chain (MLC20) of myosin, thereby regulating cell contractility. The association of MYPT1 with PP1c increases the phosphatase activity toward myosin; therefore, disrupting/dissociating this interaction may result in inhibition of the dephosphorylation of myosin. In this study, we probed how MYPT1^32–58 peptide including major PP1c interactive regions coupled with biotin and cell-penetrating TAT sequence (biotin-TAT-MYPT1) may influence MP activity. Biotin-TAT-MYPT1 inhibited the activity of MP holoenzyme and affinity chromatography as well as surface plasmon resonance (SPR) binding studies established its stable association with PP1c. Biotin-TAT-MYPT1 competed for binding to PP1c with immobilized GST-MYPT1 in SPR assays and it partially relieved PP1c inhibition by thiophosphorylated (on Thr696 and Thr853) MYPT1. Moreover, biotin-TAT-MYPT1 dissociated PP1c from immunoprecipitated PP1c-MYPT1 complex implying its holoenzyme disrupting ability. Biotin-TAT-MYPT1 penetrated into A7r5 smooth muscle cells localized in the cytoplasm and nucleus and exerted inhibition on MP with a parallel increase in MLC20 phosphorylation. Our results imply that the biotin-TAT-MYPT1 peptide may serve as a specific MP regulatory cell-penetrating peptide as well as possibly being applicable to further development for pharmacological interventions. Full article

The control of cyclin-dependent kinase 1 (CDK1) kinase activity is crucial for cell cycle progression. Cell division cycle 6 (CDC6) inhibits this activity in embryonic mitoses, and thus regulates the timing of cell division progression. The meiotic cell cycle differs greatly from the mitotic one. Metaphase II (MII)-arrested oocytes remain in prolonged M-phase state due to the high activity of CDK1 in the presence of CytoStatic Factor (CSF). The role of CDC6 in the control of CDK1 during MII and oocyte activation remains unknown. Here, we studied the role of CDC6/CDK1 interactions in Xenopus laevis cell-free extracts arrested in MII (CSF extract) and upon calcium activation leading to meiotic-to-mitotic transition. The CSF extract allows analysis of biochemical processes based on immunodepletion of selected proteins and facilitates manipulations using addition of recombinant proteins. We show by glutathione S-transferase (GST)-CDC6 pull-down that CDC6 associates with CDK1 in CSF extract and by histone H1 kinase assay that it downregulates CDK1 activity. Thus, CDC6-dependent inhibition of CDK1 is involved in the homeostasis of the MII-arrest. Upon CSF extract activation with calcium exogenous GST-CDC6 provokes accelerated transition from MII to interphase, while the depletion of endogenous CDC6 results in a slower transition to interphase. We demonstrate this by following both the phosphorylation state of CDK1 substrate cell division cycle 27 (CDC27) and histone H1 kinase assay. Importantly, increasing doses of GST-CDC6 proportionally accelerate CDK1 inactivation showing that CDC6 controls the dynamics of MII to interphase transition in a dose-dependent manner. Thus, CDC6 is a CDK1 silencer acting upon both the MII arrest and CSF extract activation by assuring the physiological activity of CDK1 during this meiotic arrest and correct timely inactivation of this kinase during the second process. Thus, we show that CDC6 controls CDK1 not only during mitotic divisions, but also in MII-arrest and the meiotic-to-mitotic transition in Xenopus laevis cell-free extracts. This study aims to bridge that gap by investigating CDC6 function using a biochemically controlled system. Full article