Biology subjects

Takahashi, T.

Publications and source records attributed to Takahashi, T..

At least 37 records · Page 2Linked to original sources

bioRxiv2024

TRBP regulates poly(I:C)-mediated apoptotic pathway by modulating miRNA biogenesis

Interferon (IFN) response is usually triggered by the invasion of viral RNAs, and is known to suppress RNA silencing triggered by microRNAs (miRNAs). However, the detailed mechanism regulating RNA silencing during IFN response has not been elucidated. We previously reported that a virus sensor protein, LGP2, which is an upregulated protein during IFN response, interacts with an RNA-binding protein, TRBP. Since TRBP is an RNA-binding protein which enhances miRNA biogenesis in collaboration with Dicer, the interaction of TRBP with LGP2 inhibits the interaction with Dicer and suppresses the TRBP-mediated miRNA biogenesis. During Sendai virus infection, we found that the inhibition of miRNA maturation caused by TRBP depletion resulted in the induction of apoptosis. However, the global gene expression network regulated by miRNAs during viral infection is not understood. In this study, we analyzed comprehensive data from RNA-sequencing and small RNA-sequencing during the antiviral response induced by poly(I:C) transfection, and investigated how TRBP regulates expression profiles of miRNAs and the downstream mRNAs. A group of miRNAs, including miRNAs preferably bind to TRBP, were downregulated by poly(I:C) transfection. Although about 60 % of miRNAs were situated in the introns of their host genes, the expression levels of mature miRNAs were not correlated with those of host genes. Therefore, the downregulation of the expression levels of mature miRNAs was considered to be not caused by transcriptional inhibition of host genes but by the inhibition of TRBP-mediated miRNA processing. Target prediction analysis suggested that the downregulated miRNAs in the presence of TRBP but not in the absence of TRBP mainly targeted transcription factors. Furthermore, these transcription factors were included in the regulators of apoptosis- and cell cycle-related factors. Some of these regulators were also predicted to be the direct targets of downregulated miRNAs in the presence of TRBP. Thus, it was revealed that the downregulation of miRNAs in the presence of TRBP regulates the expression of apoptosis- and cell cycle-related genes either directly or indirectly through adjusting the expression levels of transcription factors. These results suggest that TRBP has a pivotal role in the determination of cell fates during viral infection regulating the miRNA-mRNA regulatory networks through modifying miRNA biogenesis.

molecular biology↗

bioRxiv2024

Blocking uncertain errors of PCR

The polymerase chain reaction (PCR) plays a central role in genetic engineering and is routinely used in various applications, from biological and medical research to the diagnosis of viral infections. PCR is an extremely sensitive method for detecting target DNA sequences, but it is substantially error-prone. In particular, the mishybridization of primers to contaminating sequences can result in false positives for virus tests. The blocker method, also called the clamping method, has been developed to suppress mishybridization errors. However, its application is limited by the requirement that the contaminating template sequence must be known in advance. Here, we demonstrate that a mixture of multiple blocker sequences effectively suppresses the amplification of contaminating sequences even in the presence of uncertainty. The blocking effect was characterized by a simple model validated by experiments. Furthermore, the modeling allowed us to minimize the errors by optimizing the blocker concentrations. The results highlighted an inherent robustness of the blocker method, in that fine-tuning of the blocker concentrations is not necessary. Our method extends the applicability of PCR and other hybridization-based techniques, including genome editing, RNA interference, and DNA nanotechnology, by improving their fidelity. SignificanceThe applications of PCR are increasing day by day, and there is a need to suppress PCR errors to improve the accuracy of PCR-based techniques and broaden their applicability. The blocker method has been developed to substantially suppress mispriming. However, the method requires prior knowledge of the contaminating sequence, which limits its applicability. We successfully demonstrate that adding a combination of multiple blocker sequences can substantially suppress PCR errors, even when we have only partial information about the contaminating sequences. We also construct a biophysical model of the blocking effect, which allows us to find the optimal blocker combinations that minimize the PCR error. Since the method targets hybridization, it is readily applicable to a wide range of biotechnologies.

bioengineering↗

bioRxiv2024

Functional reorganization of brain regions supporting non-adjacent dependency learning across the first half year of life

Pre-babbling infants can track nonadjacent dependencies (NADs) in the auditory domain. While this forms a crucial prerequisite for language acquisition, the neurodevelopmental origins of this ability remain unknown. We applied functional near- infrared spectroscopy in neonates and 6-7-month-old infants to investigate the neural substrate supporting NAD learning using tone sequences in an artificial grammar learning paradigm. Detection of NADs was indicated by left prefrontal activation in neonates while by left supramarginal gyrus (SMG), superior temporal gyrus (STG), and inferior frontal gyrus activation in 6-7-month-olds. Functional connectivity analyses further indicated that the neonate activation pattern during the test phase benefited from a brain network consisting of prefrontal regions, left SMG and STG during the rest and learning phases. These findings suggest a left-hemispheric learning-related functional brain network may emerge at birth and be strengthened by complex auditory input across the first half year of life, providing a neural basis for language acquisition.

neuroscience↗

bioRxiv2024

Discovery of anti-SARS-CoV-2 S2 protein antibody CV804 with broad-spectrum reactivity with various beta coronaviruses and analysis of its pharmacological properties in vitro and in vivo

SARS-CoV-2 pandemic alerts us that spillovers of various animal coronaviruses to human in the future may bring us enormous damages. Thus, there is a significant need of antibody-based drugs to treat patients infected with previously unseen coronaviruses.CV804 against the S2 domain of the spike protein, which is less prone to mutations. CV804 shows not only broad cross-reactivities with representative 20 animal-origin coronaviruses but also with diseases-associated human beta coronaviruses including SARS-CoV, MERS-CoV, HCoV-OC43, HCoV-HKU1 and mutant strains of SARS-CoV-2. Other than that, the main characteristics of CV804 are that it has strong antibody-dependent cellular cytotoxicity (ADCC) activity to SARS-CoV2 spike protein-expressed cells in vitro and completely lacks virus-neutralization activity. Comprehensively in animal models, CV804 suppressed disease progression by SARS-CoV-2 infection. Structural studies using HDX-MS and point mutations of recombinant spike proteins revealed that CV804 binds to a unique epitope within the highly conserved S2 domain of the spike proteins of various coronaviruses. Based on the overall data, we suggest that the non-neutralizing CV804 antibody recognizes the conformational structure of the spike protein expressed on the surface of the infected cells and weakens the viral virulence by supporting host immune cells attack through ADCC activity in vivo. CV804 epitope identified in this study is not only useful for the design of pan-corona antibody therapeutics but also to design next-generation coronavirus vaccines and antiviral drugs.

microbiology↗

bioRxiv2023

Nutritional characterisation and seasonal variation of goat forages in Central Malawi

Goat ownership is prevalent across rural Malawi and provides a vital source of income, nutrition, and food security. However, goat performance is poor, and this presents a risk to individuals and communities who depend on them. Whilst mitigating this through supplementation of persevered forages may be possible, this is a challenge due to limitations of resources and the fact that goats typically free-roam during the dry season. Nutrition is fundamental to health and productivity of any livestock enterprise, it is required to be able to deal with stresses, such as disease, and to enable growth and production. The aim of this study was to characterise the nutritional profile of naturally available forages in Malawi, including the seasonal variation in nutrition. Samples of herbaceous forages and browse were collected over a 17-month period, across four villages (30 farms/smallholders) in Central Malawi. Forages underwent nutritional analysis for crude protein, fibre fractions, and ash/organic matter and NDVI obtained from satellite imagery was used as a measure of forage availability. Forage nutrition and availability were most adequate in the wet season, with higher concentrations of crude protein and a greater availability of herbaceous plants. There were significant differences in low-digestibility fibre fractions between locations, likely due to local factors such as soil and hydrology. The fall in crude protein concentrations from the wet season to the dry season represent a seasonal nutrition-gap which may result in risks to goat health and productivity.

zoology↗

bioRxiv2023

Opportunities to improve goat production and food security in Botswana through forage nutrition and the use of supplemental feeds

Goats fulfil a central role in food security across Africa with over half of households owning or rearing goats in rural areas. However, goat performance is poor and mortality high. This study assessed the nutritional quality of commonly used feeds and proposes feed-baskets to enhance goat nutrition and health. Feeds were collected from 11 areas within the Central District of Botswana, and macronutrient analyses were conducted, including crude protein, fibre fractions, ash, and metabolizable energy (ME). Forage nutrition was compared across seasons and soil types. Additionally, seasonal supplementation trials were conducted to evaluate consumption rates of various supplements, including crop residues, pellets, Lablab purpureus, and Dichrostachys cinerea. Each supplement was provided ad libitum for a 24-hour period, and consumption rates determined. Findings revealed significant differences in nutrition among various feed sources, across seasons, and in relation to soil types (p < 0.001). Consumption rates of supplements were higher during the dry season, possibly due to reduced forage availability. Supplement consumption rates varied across supplements, with crop residues accounting for approximately 1% of dry matter intake, compared to up to 45% for pellets, 13% for L. purpureus, and 15% for D. cinerea. While wet season feed baskets exhibited higher ME values compared to dry-season feed-baskets, the relative impact of supplementation was more pronounced during the dry season. These results highlight the potential for optimizing goat diets through improved grazing and browsing management, especially during the reduced nutritional availability in the dry season.

zoology↗

bioRxiv2023

Effect of prior somatosensory electrical stimulation on the wrist in biasing human hand choice

Hand choice is an unconscious decision frequently made in daily life. A correlation has been found between the state of brain activity before target presentation and hand choice for the target, around which the hand choice probability is in equilibrium. However, whether the neural state before target presentation affects hand choice remains unknown. Therefore, this study aimed to examine whether instantaneous somatosensory electrical stimulation administered to the unilateral wrist at 0, 300, or 600 ms before the target presentation facilitates or inhibits stimulated hand choice for targets around the hand selection equilibrium point. A single electrical stimulation comprised five trains of 1 ms electrical pulses, with a 20 ms inter-pulse interval. The stimulus intensity was set at 80% of the motor threshold. Fourteen right- handed healthy adults (five females, nine males; mean age, 25.1 {+/-} 4.64 years) participated. Unilateral wrist stimulation significantly increased the probability of choosing the stimulated hand and led to a faster reaction time than bilateral wrist stimulation and no-stimulation conditions. The results suggest that prior somatosensory stimulation significantly affects the hand-choice process, effectively promoting selection of the stimulated hand. These findings highlight the potential application of this stimulation method in stroke rehabilitation to facilitate use of the paretic hand. Impact StatementPrior neural stimulation significantly affected the hand-selection process by promoting the selection of the stimulated hand, thus potentially beneficial in rehabilitating the paretic hand of patients with stroke.

neuroscience↗

bioRxiv2023

Physiological roles of endocytosis and presynaptic scaffold in vesicle replenishment at fast and slow central synapses

After exocytosis, release sites are cleared of vesicular residues to replenish with transmitter-filled vesicles. Endocytic and scaffold proteins are thought to underlie this site-clearance mechanism. However, the physiological significance of this mechanism at diverse mammalian central synapses remains unknown. Here, we tested this in a physiologically optimized condition using action potential evoked EPSCs at fast calyx synapse and relatively slow hippocampal CA1 synapse, in post-hearing mice brain slices at 37{degrees}C and in 1.3 mM [Ca2+]. Pharmacological block of endocytosis enhanced synaptic depression at the calyx synapse, whereas it attenuated synaptic facilitation at the hippocampal synapse. Block of scaffold protein activity likewise enhanced synaptic depression at the calyx but had no effect at the hippocampal synapse. At the fast calyx synapse, block of endocytosis or scaffold protein activity significantly enhanced synaptic depression as early as 10 ms after the stimulation onset. Unlike previous reports, neither endocytic blockers nor scaffold protein inhibitors prolonged the recovery from short-term depression. We conclude that the release-site clearance by endocytosis can be a universal phenomenon supporting vesicle replenishment at both fast and slow synapses, whereas the presynaptic scaffold mechanism likely plays a specialized role in vesicle replenishment predominantly at fast synapses.

neuroscience↗

bioRxiv2023

Thermospermine is an evolutionarily ancestral phytohormone required for organ development and stress responses in the basal land plant Marchantia polymorpha

Thermospermine, a structural isomer of spermine, suppresses auxin-inducible xylem differentiation, whereas spermine is implicated in stress responses in angiosperms. Thermospermine synthase ACAULIS5 (ACL5) is well conserved from algae to land plants, but its physiological function remains elusive in non-vascular plants. Here we focused on MpACL5, a gene in the liverwort Marchantia polymorpha, which rescued the dwarf phenotype of the acl5 mutant of Arabidopsis. In the Mpacl5 mutants generated by genome editing, growth of the vegetative organ, thallus, and the sexual reproductive organ, gametangiophore, was severely retarded. The mutant gametangiophore exhibited remarkable morphological defects such as short stalks, fasciation, and indeterminate growth; it was formed as a fusion of two gametangiophores and a new gametangiophore was often initiated from the old one. Furthermore, Mpacl5 was shown to be hypersensitive to heat and salt stresses. Given the absence of spermine in liverworts including M. polymorpha, these results reveal that thermospermine has a dual primordial function in organ development and stress responses in the basal land plant lineage, the latter of which may have eventually been assigned to spermine during the land plant evolution.

plant biology↗

bioRxiv2023

Neuromechanical Justification of Lower-Limb Functional Tests for a Return to Running: A Muscle Coordination Analysis

IntroductionThis study aimed to explore the shared muscle synergies between running and functional tests that are commonly used when considering the return to running (RTR) after sports-related injuries. We hypothesized that shared muscle synergies would differ among tasks, providing insights into prioritizing functional tests in the context of RTR. MethodsTen healthy male participants were recruited to perform running and 9 functional tasks and their 16 lower limb and trunk muscle activities were recorded using electromyography (EMG). Non-negative matrix factorization (NMF) was applied to the collected EMG data to explore shared muscle synergies between running and the functional tasks. We compared the percentages of shared synergies and temporal patterns between running and each functional test. ResultsAlthough all functional tests exhibited shared muscle synergies with running, the walk (75% [40%-100%]), single leg hops with 30% of maximum distance (SLH30) (60% [20%-100%]), and stepup (63% [0%-100%]) tasks displayed significantly higher percentages of shared synergies compared to other tests. However, significant differences in temporal patternss were observed between running and all functional tasks, indicating varying activation profiles of shared muscle synergies. ConclusionAlthough all functional tests shared muscle synergies with running, variations in the degree of shared synergies and temporal patterns imply that walking, SLH30, and step-up tests may be the most beneficial in predicting running behavior post-ACL injuries. However, functional tests cannot fully replicate running dynamics, suggesting the need for a careful interpretation when assessing readiness for RTR.

neuroscience↗

bioRxiv2023

The topology of interpersonal neural network in weak social ties

People often have opportunities to engage in social interactions with strangers, which have been reported to contribute to their well-being. Although strategies for social interaction between strangers differ from those between acquaintances, the differences in neural basis of social interaction have not been fully elucidated. In this study, we examined the geometrical properties of interpersonal neural networks in pairs of strangers and acquaintances during joint tapping using dual electroencephalography (EEG). Twenty-one pairs of participants performed antiphase joint tapping under four different conditions. Intra-brain synchronizations were calculated using the weighted phase lag index (wPLI) for all possible intra-brain pairs of the 29 channels (29C2 = 406), and inter-brain synchronizations were calculated using the phase locking value (PLV) for all possible inter-brain pairs of the 29 channels (29 x 29 = 841) in the theta, alpha, and beta frequency bands. Electrode pairs with larger wPLI and PLV than their surrogates were defined as the nodes (EEG channels) and edges (connections between nodes) of the neural networks. We then calculated the global efficiency, local efficiency, clustering coefficient, and modularity derived from graph theory for the combined intra- and inter-brain networks of each pair. No significant differences in the tapping phase variance were identified between the stranger and acquaintance pairs. However, in the combined intra- and inter-brain theta EEG (4-7 Hz) networks, stranger pairs showed larger local efficiency and cluster coefficients than acquaintance pairs, indicating that the two brains of stranger pairs were more densely connected. Moreover, in the beta EEG bands, the modularity of the two brains was low in the fast condition, indicating that the two brains were coupled when the task demand was high. Our results show that weak social ties promote more extensive social interactions and result in dense brain-to-brain coupling.

neuroscience↗

bioRxiv2023

The Effect of Evaluating Self's Emotions on Frontal Alpha Asymmetry

In this research to assess emotions from biometric signals, participants are asked to evaluate the emotions they subjectively experienced in order to confirm whether the assumed emotions were actually elicited. However, the evaluation of emotions is not routinely performed in daily life, and it is possible that this evaluation may alter biological signals. In fMRI studies, evaluation has been shown to activate the amygdala, which is said to be related to emotional expression. However, electroencephalography (EEG) studies do not take into consideration the effects of such evaluations, and it is unclear how these evaluations affect emotion-related brain activity observed in EEG. We hypothesized that emotion evaluations would amplify emotions and c alter Frontal Alpha Asymmetry (FAA), which has been shown to be related to emotional pleasantness and unpleasantness. We suspect this is because in order to evaluate ones emotions, one must pay attention to ones internal state, and this self-focused attention has been found to enhance the subjective emotional experience. We measured a 29-channel EEG when presented with unpleasant and highly arousing images from the International Affective Picture System (IAPS) from 40 healthy male and female participants. The results revealed that FAA was significantly lower in the condition in which participants rated their own emotions compared to the condition in which they did not. Similar to fMRI studies, this result indicates that emotion-related brain activity is amplified on an EEG. This paper provides a cautionary note regarding the use of such evaluations in EEG emotion estimation studies.

neuroscience↗

bioRxiv2023

Overexpression of NIMA-related kinase suppresses cell proliferation and tip growth in a liverwort Marchantia polymorpha

NIMA-related kinases (NEKs) regulate a series of mitotic events in fungi and animals, whereas plant NEKs regulate growth direction of cells and organs. The liverwort Marchantia polymorpha has a single functional MpNEK1 gene, whose knockout leads to twisted growth of rhizoids. MpNEK1 is also expressed in the meristem of vegetative flat organ, thallus, while its function remains unknown. Here, we generated transgenic lines for the inducible expression of MpNEK1 using an estrogen receptor mediated system. Estradiol treatment efficiently induced the accumulation of MpNEK1 mRNA and MpNEK1-Citrine fusion protein throughout plant body. Overexpression of MpNEK1 severely suppressed growth of rhizoids and thalli, eventually causing the lethality of juvenile plants. The effect of estradiol was reversible until 3 days, whereas 7-days treatment resulted in irreversible suppression of growth. This severe effect was observed even at the nanomolar level of estradiol. EdU staining and microtubule imaging clearly indicated the suppression of cell proliferation by estradiol-induced MpNEK1. Unexpectedly, the overexpression of kinase-deficient MpNEK1 also suppressed thallus growth and rhizoid formation, despite their slightly mild effect than the full length MpNEK1, indicating phosphorylation-independent mechanism of growth suppression. In conclusion, overexpression of MpNEK1 suppresses cell division and elongation, leading to growth cessation and lethality. Our results imply that the expression of MpNEK1 is tightly regulated and plant NEKs might control cell division as in fungi and animals.

plant biology↗

bioRxiv2022

Golgi retention and oncogenic KIT signaling via PLCγ2-PKD2-PI4KIIIβ activation in GIST cells

Most gastrointestinal stromal tumors (GISTs) develop due to gain-of-function mutations in the tyrosine kinase, KIT. We recently showed that mutant KIT mislocalizes to the Golgi area and initiates uncontrolled signaling. However, the molecular mechanisms underlying its Golgi retention remain unknown. Here, we show that protein kinase D2 (PKD2) is activated by the mutant, which causes KITs Golgi retention. In PKD2-inhibited cells, KIT migrates from the Golgi region to lysosomes and subsequently undergoes degradation. Importantly, delocalized KIT is unable to trigger downstream activation. In the Golgi area, KIT activates the PKD2-phosphatidylinositol 4-kinaseIII{beta} (PKD2-PI4KIII{beta}) pathway through phospholipase {gamma}2 (PLC{gamma}2) to generate a PI4P-rich membrane domain, where the AP1-GGA1 complex is aberrantly recruited. Disruption of any factors in this cascade results in KIT release from the Golgi region, indicating that these PKD2-related pathways are responsible for the Golgi retention of KIT. Our findings unveil the molecular mechanisms underlying KIT mislocalization and provide evidence for a new strategy for inhibition of oncogenic signaling.

cancer biology↗

bioRxiv2022

RNA processing/modifying enzymes play key roles in the response to thermospermine in Arabidopsis thaliana

Thermospermine acts in negative regulation of xylem differentiation through enhancing mRNA translation of the members of the SAC51 gene family in Arabidopsis. These mRNAs contain conserved upstream open-reading-frames (uORFs) that are inhibitory to the main ORF translation. To address the mode of action of thermospermine in this process, we have isolated mutants that are insensitive to thermospermine, named its. We show here that four genes responsible for the mutants, its1 to its4, encode a homologue of SPOUT RNA methyl transferase, an rRNA pseudouridine synthase CBF5/NAP57, a putative spliceosome disassembly factor STIPL1/NTR1, and a plant-specific RNA-binding protein PHIP1, respectively. While these mutants except its1 are almost normal in appearance, they enhance the dwarf phenotype of a mutant of ACL5 defective in thermospermine biosynthesis, resulting in tiny-sized plants reminiscent of a double knockout of ACL5 and SACL3, a member of the SAC51 family. We confirmed that the GUS reporter activity from the SAC51 5-GUS fusion transcript was severely reduced in all of these mutants. These results unveil the importance of RNA processing and modification for the translation of transcripts containing regulatory uORFs.

plant biology↗

bioRxiv2022

Strategy for selective acquisition of transgenic marmosets using the piggyBac transposon system

Transgenic (Tg) nonhuman primates (NHPs) are excellent potential models due to their physiological similarities with humans. In NHPs, the lentiviral vector is the only available transgenesis method, but transgene sizes are limited. The piggyBac (PB) transposon system is a DNA element sandwiched between two terminal inverted repeats (TRs) that allows transposases to introduce larger genes than lentiviral vectors. However, embryos with integrated transgenes are difficult to distinguish using this system. To address this issue, we placed the Enhanced Green Fluorescent Protein (EGFP) gene inside the TRs and the humanized kusabira orange 1 (hKO1) fluorescent protein gene with the proteolysis-promoting sequence d2PEST outside the TRs to create an indicator for gene integration using fluorescent protein expression. In HEK293T cells, hKO1 and EGFP co-expression after transfection gradually changed to decay of hKO1 expression and enhancement of EGFP expression by transposase within five days. In mouse and marmoset embryos, hKO1 expression was attenuated, while only EGFP expression was observed at the blastocyst stage. All offspring obtained through embryo transfer of EGFP-expressing embryos were Tg; thus, we established a new PB system capable of determining Tg embryos and successfully produced Tg marmosets, for the first time, in NHPs.

molecular biology↗

bioRxiv2022

Plant specific armadillo repeat kinesin directs organelle transport and microtubule convergence to promote tip growth

Tip growth is essential for plant growth and reproduction. However, it remains elusive how highly polarized tip-growing cells coordinate the organization of intracellular components. Here, we show that plant-specific armadillo-repeat containing kinesin, MpARK, mediates organelle transport and microtubule convergence in tip growing rhizoids in the early diverging land plant Marchantia polymorpha. MpARK is required for anterograde transport of organelles to maintain their position. Furthermore, MpARK participates in the formation of microtubule foci at the rhizoid apex to stabilize growth direction. MpARK-dependent rhizoid growth is essential for plant anchorage and soil holding capacity. Thus, ARK might be a principal organelle transporter and intracellular organizer in the primitive rooting cells for the adaptation to terrestrial environments. Our findings suggest that ARK is functionally comparable to canonical anterograde kinesins in animals and fungi.

cell biology↗

bioRxiv2022

The termination of UHRF1-dependent PAF15 ubiquitin signaling is regulated by USP7 and ATAD5

UHRF1-dependent ubiquitin signaling plays an integral role in the regulation of maintenance DNA methylation. UHRF1 catalyzes transient dual mono-ubiquitylation of PAF15 (PAF15Ub2), which regulates the localization and activation of DNMT1 at DNA methylation sites during DNA replication. Although the initiation of UHRF1-mediated PAF15 ubiquitin signaling has been relatively well characterized, mechanisms underlying its termination and how they are coordinated with the completion of maintenance DNA methylation have not yet been clarified. This study shows that deubiquitylation by USP7 and unloading by ATAD5 (ELG1 in yeast) are pivotal processes for the removal of PAF15 from chromatin. On replicating chromatin, USP7 specifically interacts with PAF15Ub2 in a complex with DNMT1. USP7 depletion or inhibition of the interaction between USP7 and PAF15 results in abnormal accumulation of PAF15Ub2 on chromatin. Furthermore, we also find that the non-ubiquitylated form of PAF15 (PAF15Ub0) is removed from chromatin in an ATAD5-dependent manner. PAF15Ub2 was retained at high levels on chromatin when the catalytic activity of DNMT1 was inhibited, suggesting that the completion of maintenance DNA methylation is essential for termination of UHRF1-mediated ubiquitin signaling. This finding provides a molecular understanding of how the maintenance DNA methylation machinery is disassembled at the end of the S phase.

molecular biology↗