Biology subjects

Hill, R.

Publications and source records attributed to Hill, R..

At least 19 recordsLinked to original sources

bioRxiv2026

Mechanistically distinct BER processes are essential for the tolerance of exogenous 5hm-dC and enzymatic oxidative demethylation

Epigenetic information is transmitted through covalent DNA modifications that regulate chromatin structure and gene expression. 5-Hydroxymethylcytosine (5hmC) is a key epigenetic intermediate generated during TET-mediated oxidative demethylation of 5-methylcytosine. Whether 5hmC itself is intrinsically genotoxic remains unclear. Here, we combine genome-wide CRISPR loss-of-function screens, isogenic knockouts and mass spectrometry to systematically compare the cellular consequences of exogenous 5-hydroxymethyl-2'-cytidine (5hm-dC) exposure and endogenous oxidative demethylation in mammalian cells. We find that exogenous 5hm-dC causes toxicity, mediated by deamination to 5-hydroxymethyl-2'-deoxyuridine (5hm-dU), followed by excision by the glycosylase SMUG1, which generates base excision repair (BER) intermediates that compromise cell viability. In contrast, toxicity associated with enzymatic oxidative demethylation is primarily driven by TDG-dependent excision of oxidized methylcytosine derivatives. Despite these distinct initiating events, both genotoxins converge on a critical requirement for DNA polymerase {beta} (POL{beta}), indicating that efficient BER completion is essential to mitigate cytotoxic repair intermediates.

molecular biology↗

bioRxiv2026

The impact of long-read sequencing on fungal genome assemblies: progress and disparity

Fungal genomics has expanded rapidly over the past 30 years, and recently the pace and breath has further quickened for many taxa, although many taxonomic gaps persist. With three decades of rapid growth, fungal genomics now merits a re-examination of its history, progress, and unresolved taxonomic gaps. Here, we review the development of fungal genomics from early efforts such as the Fungal Genome Initiative to current progress driven by third-generation long-read sequencing. We have compiled and summarised publicly available fungal genomes to highlight trends in assembly quality, adoption of long-read technologies, and taxonomic representation. Notably, substantial phylogenetic gaps remain, particularly outside Dikarya, and significant challenges persist for unculturable taxa. This review identifies priorities for the fungal community, including: (1) coordinated efforts to close major taxonomic gaps across the fungal tree of life; (2) improved repository metrics to facilitate identification of high-quality assemblies; and (3) improved and standardised genome annotation which is lacking for most assemblies. Together, these steps will support the development of reliable genomic resources that capture the full breadth of diversity across the fungal kingdom, generating foundational data for comparative genomics, evolutionary biology, functional studies, genetic studies and applied research.

genomics↗

bioRxiv2026

Transcriptional signatures underlying divergent lifestyles of endophytic and pathogenic fungi in early colonisation of wheat roots

Wheat take-all is a root disease which devastates crop yields, caused by the ascomycete fungus Gaeumannomyces tritici. The closely related root endophyte, G. hyphopodioides, has been found to induce local host defence responses which confer protection against take-all and reduce disease severity. Chancellor et al. (2024) investigated host transcriptional response to early colonisation by each of these two fungi. Using this RNA-seq dataset in conjunction with newly available Gaeumannomyces reference genomes, we have completed the picture by characterising the fungal transcriptional activity underpinning these different lifestyles. Even at early time points, their transcriptional profiles differ: G. hyphopodioides shows signs of transcriptional reprogramming between 4 and 5 days post inoculation (dpi), mirroring the wheat response, whereas G. tritici expression varied very little between these two time points despite progressing into the vasculature, instead exhibiting a stealthy expression profile dominated by gene downregulation at earlier time points. Moreover, GO term enrichment in this study identified a stress-response unique to G. hyphopodioides, which may explain the formation of its subepidermal vesicles (SEVs), putative resting structures that are a key difference between the pathogen and non-pathogen, alongside upregulation of many putative effectors and CAZymes. The enrichment of a key lignin-degrading CAZyme may contribute to the lack of stress-response identified in G. tritici, allowing fungal hyphae to overcome localised host lignification. These findings highlight the transcriptional basis of colonisation differences and are a step towards understanding how closely related fungi with different lifestyles modulate their interactions within a common host and tissue.

evolutionary biology↗

bioRxiv2026

Phylogenomic analyses of the Austral podocarps (Podocarpus: Podocarpaceae) reveals unlikely hybrid ancestry of a New Zealand species

Hybridisation is ubiquitous amongst plants and has important evolutionary consequences ranging from the collapse of distinct lineages through to the generation of new species. Here, we develop a phylogenetic hypothesis for the Austral podocarps (Podocarpus), a monophyletic group of six species distributed in Tasmania, mainland Australia, New Zealand and New Caledonia, and identify a putative hybrid lineage. Using a targeted capture approach to generate DNA sequence data, we find discordance between nuclear and plastid derived phylogenetic estimates and in particular, the relationships of the New Zealand species Podocarpus nivalis and Australian P. lawrencei are significantly discordant. Species network analyses largely resolve this incongruence and indicate that P. nivalis is a hybrid lineage, with P. laetus (New Zealand) and P. lawrencei as parents. We hypothesise that P. nivalis has arisen following trans-Tasman dispersal of P. lawrencei, and shows eco-geographic divergence from P. laetus, which could facilitate reproductive isolation. We suggest that introgression from P. laetus to colonising P. lawrencei could significantly reduce founder effects while cold tolerance inherited from P. lawrencei has enabled P. nivalis to occupy alpine environments. Our findings highlight the importance of reticulate evolution in Southern Hemisphere conifers and demonstrate the value of phylogenomic network approaches for resolving recent and complex radiations.

evolutionary biology↗

bioRxiv2025

Fast calcium-dependent reorientation of motile cilia basal bodies in the simple metazoan, Trichoplax.

Ciliary-based animal locomotion relies on the spatially coordinated beating of motile cilia to displace the body in liquid environments or across solid surfaces. The orientation of ciliary beating largely depends on the rotational polarity of the basal body, which in most animals is fixed and controlled by cues linked to the main body axes. The small marine animal Trichoplax exploits the beating of motile cilia in its lower epithelium to crawl on substrates. However, Trichoplax lacks defined body axes and exhibits rapid changes in body shape and direction of movement, thus raising the question of what controls the orientation and reorientation of ciliary beating. We show here that the basal bodies of the cilia in the lower epithelium of Trichoplax are oriented along the direction of the animal movements, and that they change their orientation throughout the lower epithelium in a few seconds time when the animal modifies its shape or changes direction following external mechanical stimuli. We also show that Ca2+ is required for fast basal body reorientation. Such rapid ciliary basal body reorientation has never been observed in metazoans before. Thus, our results identify a previously undescribed mechanism underlying directional motility in a metazoan and shed light on its subcellular determinants, bridging the scale between intra-cellular ciliary machinery and animal movement.

cell biology↗

bioRxiv2025

Generation of two Betacellulin CRISPR-Cas9 knockout hiPSC lines to study affected EGF system paradigm in Schizophrenia

Several members of the epidermal growth factor (EGF) family have been implicated in the biology of schizophrenia (Ketharanathan et al., 2024). The EGF-related ligand, Betacellulin (BTC), plays an important role in the proliferation and differentiation of neural stem cells and our group found markedly reduced BTC levels in patients with schizophrenia. Nevertheless, the interplay of affected BTC and its participation in neural specification and neurodevelopment remains elusive. We generated Knockout (KO) - BTC clones from an existing hiPSC line through CRISPR/Cas9-mediated modification. Furthermore, we validated BTC-KO through genotyping/sequencing, FACS and Western Blot. Finally, we demonstrated trilineage differentiation potential in vitro.

neuroscience↗

bioRxiv2025

Leveraging existing data to maximise quality and consistency across gene model annotations: a Fusarium pan-annotation

Comparative genomics analyses are frequently used to inform our understanding of how organisms have evolved and how genetics contributes to phenotypic traits. Thanks to the considerable growth in the number of sequenced and assembled genomes, there is increasingly abundant data with which to perform such analyses. However, the appropriate use of genomic data can be highly dependent on genome annotation. Genome annotation is the critical step for providing biological context to genome sequences, but due to the complexity of the task it remains a major bottleneck in analyses. Different methods, and a lack of widely accepted standards, can also result in a great diversity of completeness and accuracy across genome annotations. Accordingly, comparative genomics analyses are susceptible to errors which can be misinterpreted as biological variation, yet efforts to revise and update existing annotations have not kept pace with advances in technology and expanding data resources. We have developed a workflow to utilise existing genome annotations alongside de novo gene predictions to improve both the collective consistency and individual quality of genome annotations of a closely related group of genomes. In this work we apply this new workflow on a dataset of 82 genomes from the economically, ecologically and clinically important fungal genus Fusarium. We show that both individual as well as collective annotation quality can be improved. The development of reannotation approaches such as we present here will be essential if we are to capitalise on the huge investment that has gone into generating existing genome data. The Fusarium pan-annotation is available from Zenodo at https://zenodo.org/doi/10.5281/zenodo.13829922. Workflow code and sample commands are available from https://github.com/EI-CoreBioinformatics/FusariumPanAnno. KEY POINTSO_LIComparative genomics is a fundamental approach to understand the contributions of genetic features to biological questions. C_LIO_LITo take advantage of existing data, most comparative genomics studies compare gene models produced using a variety of annotation methodologies, which introduces computational bias that can be misinterpreted as biological signal. C_LIO_LIWe present a bioinformatics workflow to improve consistency across a set of gene model annotations and minimise computational bias for downstream comparative genomics analyses. C_LIO_LIReannotation of a previously published Fusarium dataset reaffirms the finding that comparing annotations generated from a mix of methodologies can underestimate core genes, overestimate taxon-specific genes and confound patterns of gene presence/absence. C_LI

genomics↗

bioRxiv2024

Auto-antibodies against interferons are common in people living with chronic hepatitis B virus infection and associate with PegIFN non-response

Background and aimsType one (T1) and three interferons (T3IFNs) are implicated in Chronic Hepatitis B (CHB) immunopathogenesis. IFN remains the only licenced immune modulating therapy for CHB. We measured the prevalence of auto-antibodies (auto-Abs) against T1 and T3IFNs to examine the hypothesis that they impact Hepatitis B Virus (HBV) control and treatment response, as highlighted by COVID-19. MethodsOur multi-centre retrospective longitudinal study accessed two CHB cohorts, auto-Ab levels and neutralisation status were measured against T1IFN and T3IFN. Associations were tested against HBV clinical parameters. ResultsOverall, 11.9% (33/276) of CHB patients had any detectable anti-IFN auto-Abs and 9.8% (27/276) anti-T3IFN auto-Abs, with high incidence of PegIFN-induced de novo auto-Ab (25.7%, 9/35). However, only a minority of auto-Ab-positive sera demonstrated neutralisation in vitro (3/33, 9.1%). Auto-Ab-positivity correlated with higher median HBsAg levels (p=0.0024). All individuals with detectable auto-Ab were PegIFN non-responders including those without auto-Ab against IFN specifically. ConclusionsNon-neutralising anti-IFN auto-Abs are common in CHB and associate with higher median HBsAg levels. Further prospective study of anti-cytokine auto-Ab in CHB are required to characterise association with long-term outcomes. Impact and implicationsHBV and PegIFN individually may induce broad auto-reactivity associated with dysregulated anti-viral immune responses. Auto-Ab screening pre-PegIFN and other immunotherapies may have a critical role in stratifying patient selection.

immunology↗

bioRxiv2024

Starship giant transposable elements cluster by host taxonomy using kmer-based phylogenetics

Starships are a recently established superfamily of giant cargo-mobilising transposable elements in the fungal subphylum Pezizomyotina (phylum Ascomycota). To date, Starship elements have been identified up to [~]700 Kbp in length and carrying hundreds of accessory genes, which can confer both beneficial and deleterious traits to the host genome. Classification of Starship elements has been centred on the tyrosine recombinase gene that mobilises the element, termed the captain. We contribute a new perspective to Starship classification by using an alignment-free kmer-based phylogenetic tree building method, which can infer relationships between elements in their entirety, including both active and degraded elements and irrespective of high variability in element length and cargo content. In doing so we found that relationships between entire Starships differed from those inferred from captain genes and revealed patterns of element relatedness corresponding to host taxonomy. Using Starships from Gaeumannomyces species as a case study, we found that kmer-based relationships correspond with similarity of cargo gene content. Our results suggest that Starship-mediated horizontal transfer events are frequent between species within the same genus but are less prevalent across larger host evolutionary distances. This novel application of a kmer-based phylogenetics approach overcomes the issue of how to represent and compare highly variable Starships elements as a whole, and in effect shifts the perspective from a captain to a cargo-centred concept of Starship identity. SUMMARYWe applied a kmer-based phylogenetic classification approach to giant Starship cargo-mobilising elements from species across the Pezizomycotina (Ascomycota, Fungi). We found Starship elements to frequently cluster according to host taxonomy, suggesting horizontal transfer of elements is less common across larger evolutionary distances. Kmer-based phylogenetics approaches show promise for both element classification and to inform our understanding of the evolution of Starships and other giant cargo-mobilising elements.

evolutionary biology↗

bioRxiv2024

Long Amplicon Nanopore Sequencing for Dual-Typing RdRp and VP1 Genes of Norovirus Genogroups I and II in Wastewater

Noroviruses (NoV) are the leading cause of non-bacterial gastroenteritis across the globe with societal costs of US$60.3 billion per annum. Development of a long amplicon nanopore-based method for dual-typing the RNA-dependent RNA polymerase (RdRp) and major structural protein (VP1) regions from a single RNA fragment could improve existing norovirus typing methods. Its application to wastewater-based epidemiology (WBE) and environmental testing could enable the discovery of novel types and improve tracking throughout the population and into aquaculture and recreational water settings. Here, we develop and optimise such a method for wastewater as the sample matrix. Reverse transcription (RT), PCR and library pooling were optimised and a consensus-based bioinformatics pipeline was developed. Inhibitor removal and LunaScript(R) RT gave robust amplification of the {approx}1000 bp RdRP+VP1 amplicon. Platinum Taq polymerase showed good sensitivity and reduced levels non-specific amplification (NSA) when compared to other polymerases. Optimised PCR annealing temperatures significantly reduced NSA (51.3% and 42.4% for GI and GII), increased yield (86.5% for GII) and increased taxa richness (57.7%) for GII. Analysis of three NoV positive faecal samples showed 100% nucleotide similarity with Sanger sequencing. Eight GI genotypes, 11 polymerase types (p-types) and 13 combinations were detected in wastewater along with 4 GII genotypes, 4 p-types and 8 combinations; highlighting the diversity of norovirus taxa present in wastewater in England. The most common genotypes detected in clinical samples were all detected in wastewater while we also commonly detected several GI genotypes not reported in the clinical data. Application of this method into a WBE scheme, therefore, may allow for more accurate measurement of norovirus diversity within the population.

molecular biology↗

bioRxiv2024

Evolutionary genomics reveals variation in structure and genetic content implicated in virulence and lifestyle in the genus Gaeumannomyces

Gaeumannomyces tritici is responsible for take-all disease, one of the most important wheat root threats worldwide. High-quality annotated genome resources are sorely lacking for this pathogen, as well as for the closely related antagonist and potential wheat take-all biocontrol agent, G. hyphopodioides. As such, we know very little about the genetic basis of the interactions in this host-pathogen-antagonist system. Using PacBio HiFi sequencing technology we have generated nine near-complete assemblies, including two different virulence lineages for G. tritici and the first assemblies for G. hyphopodioides and G. avenae (oat take-all). Genomic signatures support the presence of two distinct virulence lineages in G. tritici (types A and B), with A strains potentially employing a mechanism to prevent gene copy-number expansions. The CAZyme repertoire was highly conserved across Gaeumannomyces, while candidate secreted effector proteins and biosynthetic gene clusters showed more variability and may distinguish pathogenic and non-pathogenic lineages. A transition from self-sterility (heterothallism) to self-fertility (homothallism) may also be a key innovation implicated in lifestyle. We did not find evidence for transposable element and effector gene compartmentalisation in the genus, however the presence of Starship giant transposable elements likely contributes to genomic plasticity in the genus. Our results depict Gaeumannomyces as an ideal system to explore interactions within the rhizosphere, the nuances of intraspecific virulence, interspecific antagonism, and fungal lifestyle evolution. The foundational genomic resources provided here will enable the development of diagnostics and surveillance of understudied but agriculturally important fungal pathogens.

evolutionary biology↗

bioRxiv2022

Mouse behaviour on the trial-unique non-matching-to-location (TUNL) touchscreen task reflects a mixture of distinct working memory codes and response biases

The trial-unique non-matching to location (TUNL) touchscreen task shows promise as a translational assay of working memory deficits in disorders including autism, ADHD, and schizophrenia. Although it is commonly assumed that the TUNL task predominantly measures spatial working memory in rodents, this proposition has not previously been tested. In this project, we used computational modelling of behaviour from mice performing the TUNL task (total N = 163 mice across three datasets; 158,843 total trials). Contrary to common assumptions, behaviour on the TUNL task did not exclusively reflect spatial working memory. Instead, choice behaviour was explained as a mixture of both retrospective (spatial) working memory and prospective working memory for an intended behavioural response, as well as animal-specific response biases. We suggest that these findings can be understood within a resource-rational cognitive framework, and propose several task-design principles that we predict will maximise spatial working memory and minimise alternative behavioural strategies.

animal behavior and cognition↗

bioRxiv2022

Medulloblastoma Group 3 and 4 Tumors Comprise a Clinically and Biologically Significant Expression Continuum Reflecting Human Cerebellar Development

Medulloblastoma is currently sub-classified into distinct DNA methylation subgroups/subtypes with particular clinico-molecular features. Using RNA-seq in large well annotated cohorts of medulloblastoma we show that transcriptionally Group3 and Group4 medulloblastomas exist not as discrete types but as intermediates on a bipolar continuum between archetypal Group3 and Group4 entities. Continuum position is prognostic, reflects propensity for specific DNA copy-number changes, key switches in isoform/enhancer usage and RNA-editing. Examining scRNA-seq profiles we show intra-tumoral transcriptional heterogeneity along the continuum is limited in a subtype-dependent manner. By integrating with a human scRNA-seq reference atlas we show this continuum is mirrored by an equivalent continuum of transcriptional cell types in early fetal cerebellar development. We identify unique developmental niches for all four major subgroups and link each to a common developmental antecedent. Our findings show a transcriptional continuum arising from oncogenic disruption of highly specific fetal cerebellar cell types, linked to almost every aspect of Group3/Group4 molecular biology and clinico-pathology.

cancer biology↗

bioRxiv2022

Model selection in occupancy models: inference versus prediction

Occupancy models are a vital tool for applied ecologists studying the patterns and drivers of species occurrence, but their use requires a method for selecting between models with different sets of occupancy and detection covariates. The information-theoretic approach, which employs information criteria such as Akaikes Information Criterion (AIC) is arguably the most popular approach for model selection in ecology and is often used for selecting occupancy models. However, the information-theoretic approach risks selecting models which produce inaccurate parameter estimates, due to a phenomenon called collider bias. Using simulations, we investigated the consequences of collider bias (using an illustrative example called M-bias) in the occupancy and detection processes of an occupancy model, and explored the implications for model selection using AIC and a common alternative, the Schwarz Criterion (or Bayesian Information Criterion, BIC). We found that when M-bias was present in the occupancy process, AIC and BIC selected models which inaccurately estimated the effect of the focal occupancy covariate, while simultaneously producing more accurate predictions of the site-level occupancy probability. In contrast, M-bias in the detection process did not impact the focal estimate; all models made accurate inferences, while the site-level predictions of the AIC/BIC-best model were slightly more accurate. Our results demonstrate that information criteria can be used to select occupancy covariates if the sole purpose of the model is prediction, but must be treated with more caution if the purpose is to understand how environmental variables affect occupancy. By contrast, detection covariates can usually be selected using information criteria regardless of the models purpose. These findings illustrate the importance of distinguishing between the tasks of parameter inference and prediction in ecological modelling. Furthermore, our results underline concerns about the use of information criteria to compare different biological hypotheses in observational studies. Open Research StatementCode to fully reproduce our simulations and analyses is available at: https://zenodo.org/badge/latestdoi/462801230

ecology↗

bioRxiv2020

In vitro hydroxyapatite formation of a TTCP and DCPA based dentine desensitiser

The objective of this study was to characterise apatite formation ability of a tetra calcium phosphate (TTCP) and dicalcium phosphate anhydrous (DCPA) based desensitising product, Teethmate (TM). Methods: Cylindrical samples of TM were produced and immersed in TRIS buffer and artificial saliva (AS) for selected time periods. This was followed by characterisation of the phases formed using 31P Magic Angle - Nuclear Magnetic Resonance Spectroscopy, X-ray powder diffraction and Fourier-transform infrared spectroscopy. Dentine discs treated with the material were analysed by scanning electron microscopy (SEM) to evaluate crystal shape and tubules occlusion. Results: SEM analysis showed the formation of plate-like crystals occluding the dentinal tubules. Solid-state characterisation analyses showed that TM cement samples immersed in TRIS-buffer (pH: 7.3) formed hydroxyapatite phases within approximately six hours of immersion. HA formation was less rapid for the samples stored in AS (pH: 6.5). Conclusions: This study shows that a TTCP-DCPA based desensitising product (TM) converts to HA in vitro and provides an effective option for the treatment of dentine hypersensitivity.

biophysics↗

bioRxiv2020

In vitro hydroxyapatite formation of a TTCP and DCPA based dentine desensitiser

The objective of this study was to characterise apatite formation ability of a tetra calcium phosphate (TTCP) and dicalcium phosphate anhydrous (DCPA) based desensitising product, TeethmateTM (TM). Methods: Cylindrical samples of TM were produced and immersed in TRIS buffer and artificial saliva (AS) for selected time periods. This was followed by characterisation of the phases formed using 31P Magic Angle - Nuclear Magnetic Resonance Spectroscopy, X-ray powder diffraction and Fourier-transform infrared spectroscopy. Dentine discs treated with the material were analysed by scanning electron microscopy (SEM) to evaluate crystal shape and tubules occlusion. Results: SEM analysis showed the formation of plate-like crystals occluding the dentinal tubules. Solid-state characterisation analyses showed that TM cement samples immersed in TRIS-buffer (pH: 7.3) formed hydroxyapatite phases within approximately six hours of immersion. HA formation was less rapid for the samples stored in AS (pH: 6.5). Conclusions: This study shows that a TTCP-DCPA based desensitising product (TM) converts to HA in vitro and provides an effective option for the treatment of dentine hypersensitivity.

biophysics↗

bioRxiv2019

A Highly Conserved Shh Enhancer Coordinates Hypothalamic and Craniofacial Development

Enhancers that are conserved deep in evolutionary time regulate characteristics held in common across taxonomic classes. Here, deletion of the highly conserved Shh enhancer SBE2 (Shh brain enhancer 2) markedly reduced Shh expression within the embryonic brain specifically in the rostral diencephalon; however, no abnormal anatomical phenotype was observed due to compensatory low levels of expression mediated by a second enhancer. In contrast, a further reduction of Shh levels, achieved by crossing the SBE2 deletion with the Shh null allele, disrupted brain and craniofacial tissue; thus, linking SBE2 regulated Shh expression to multiple defects and further enabling the study of the effects of differing levels of Shh on embryogenesis. Development of the hypothalamus, derived from the rostral diencephalon, was disrupted along both the anterior-posterior (AP) and the dorsal-ventral (DV) axes. Expression of DV patterning genes and subsequent neuronal population induction were particularly sensitive to Shh expression levels, demonstrating a novel morphogenic context for Shh. The role of SBE2, which is highlighted by DV gene expression, is to step-up expression of Shh above the minimal activity of the second enhancer, ensuring the necessary levels of Shh in a regional-specific manner. We also show that low Shh levels in the diencephalon disrupted neighbouring craniofacial development, including mediolateral patterning of the bones along the cranial floor and viscerocranium. Thus, SBE2 contributes to hypothalamic morphogenesis and ensures there is coordination with the formation of the adjacent midline cranial bones that subsequently protects the neural tissue.

developmental biology↗

bioRxiv2019

JMJD6 Cleaves MePCE to Release P-TEFb

More than 30% of genes in higher eukaryotes are regulated by promoter-proximal pausing of RNA polymerase II (Pol II). Phosphorylation of Pol II-CTD by positive transcription elongation factor (P-TEFb) is a necessary precursor event that enables productive transcription elongation. The exact mechanism on how the sequestered P-TEFb is released from the 7SK snRNP complex and recruited to Pol II-CTD remains unknown. In this report, we reveal methylphosphate capping enzyme (MePCE), a core component of the 7SK snRNP complex, as the cognate substrate for Jumonji domain-containing 6 (JMJD6)s novel proteolytic function. Our evidences consist of a crystal structure of JMJD6 bound to methyl-arginine, enzymatic assays of JMJD6 cleaving MePCE in vivo and in vitro, binding assays, and downstream effects of Jmjd6 knockout and overexpression on Pol II-CTD phosphorylation. We propose that JMJD6 assists bromodomain containing 4 (BRD4) to recruit P-TEFb to Pol II-CTD by disrupting the 7SK snRNP complex.

biochemistry↗