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The Integrator complex is a multisubunit complex that participates in the processing of small nuclear RNA molecules in eukaryotic cells by cleaving the 3’ end. In protein-coding genes, Integrator is a key regulator of promoter-proximal pausing, release, and recruitment of RNA polymerase II. Research on Integrator has revealed its critical role in the regulation of gene expression and RNA processing. Dysregulation of the Integrator complex has been implicated in a variety of human diseases including cancer and developmental disorders. Therefore, understanding the structure and function of the Integrator complex is critical to uncovering the mechanisms of gene expression and developing potential therapeutic strategies for related diseases. [ABSTRACT FROM AUTHOR]

Mammalian genomes produce an abundance of short RNA. This is, to a large extent, due to the genome-wide and spurious activity of RNA polymerase II (RNAPII). However, it is also because the vast majority of initiating RNAPII, regardless of the transcribed DNA unit, terminates within a ∼3-kb early "pausing zone." Given that the resultant RNAs constitute both functional and non-functional species, their proper sorting is critical. One way to think about such quality control (QC) is that transcripts, from their first emergence, are relentlessly targeted by decay factors, which may only be avoided by engaging protective processing pathways. In a molecular materialization of this concept, recent progress has found that both "destructive" and "productive" RNA effectors assemble at the 5′ end of capped RNA, orchestrated by the essential arsenite resistance protein 2 (ARS2) protein. Based on this principle, we here discuss early QC mechanisms and how these might sort short RNAs to their final fates. [Display omitted] The majority of initiating RNA polymerase II (RNAPII) terminates early, generating an abundance of short RNAs that require sorting into productive or destructive pathways. Here, Garland and Jensen explore early quality-control mechanisms that govern the fate of short RNAs, preventing the accumulation of non-functional transcripts and ensuring proper RNA processing. [ABSTRACT FROM AUTHOR]

Endogenous retroviruses (ERVs) are remnants of ancient parasitic infections and comprise sizable portions of most genomes. Although epigenetic mechanisms silence most ERVs by generating a repressive environment that prevents their expression (heterochromatin), little is known about mechanisms silencing ERVs residing in open regions of the genome (euchromatin). This is particularly important during embryonic development, where induction and repression of distinct classes of ERVs occur in short temporal windows. Here, we demonstrate that transcription-associated RNA degradation by the nuclear RNA exosome and Integrator is a regulatory mechanism that controls the productive transcription of most genes and many ERVs involved in preimplantation development. Disrupting nuclear RNA catabolism promotes dedifferentiation to a totipotent-like state characterized by defects in RNAPII elongation and decreased expression of long genes (gene-length asymmetry). Our results indicate that RNA catabolism is a core regulatory module of gene networks that safeguards RNAPII activity, ERV expression, cell identity, and developmental potency.

Repressor element-1 silencing transcription factor (REST) is required for the formation of mature neurons. REST dysregulation underlies a key mechanism of neurodegeneration associated with neurological disorders. However, the mechanisms leading to alterations of REST-mediated silencing of key neurogenesis genes are not known. Here, we show that BRCA1 Associated ATM Activator 1 (BRAT1), a gene linked to neurodegenerative diseases, is required for the activation of REST-responsive genes during neuronal differentiation. We find that INTS11 and INTS9 subunits of Integrator complex interact with BRAT1 as a distinct trimeric complex to activate critical neuronal genes during differentiation. BRAT1 depletion results in persistence of REST residence on critical neuronal genes disrupting the differentiation of NT2 cells into astrocytes and neuronal cells. We identified BRAT1 and INTS11 co-occupying the promoter region of these genes and pinpoint a role for BRAT1 in recruiting INTS11 to their promoters. Disease-causing mutations in BRAT1 diminish its association with INTS11/INTS9, linking the manifestation of disease phenotypes with a defect in transcriptional activation of key neuronal genes by BRAT1/INTS11/INTS9 complex. Finally, loss of Brat1 in mouse embryonic stem cells leads to a defect in neuronal differentiation assay. Importantly, while reconstitution with wild-type BRAT1 restores neuronal differentiation, the addition of a BRAT1 mutant is unable to associate with INTS11/INTS9 and fails to rescue the neuronal phenotype. Taken together, our study highlights the importance of BRAT1 association with INTS11 and INTS9 in the development of the nervous system. [ABSTRACT FROM AUTHOR]

In the course of this study, an efficient implementation of a high gain and low power two-stage operational amplifier using indirect feedback frequency compensation based on CNTFETs. HSPICE software was used to develop and simulate CNTFETs. The op-amps were designed using 0.9 V input supply voltage. The proposed structures were formed either using CNTFETs only known as pure CNTFET-IFFC-2SOA or hybrid structures consisting a mix of both CNTFETs and conventional CMOS named as PCNTFET-NMOS-IFFC-2SOA and NCNTFET-PMOS-2SOA. The comparative investigation revealed that CNT-based structures performed significantly better than the traditional CMOS-based devices. In the instance of CNTFET-IFFC-2SOA, there was a considerable improvement in DC gain, power dissipation, phase margin, and CMRR. The DC Gain increased by 140.92%, the CMRR increased by 32.94%, and the phase margin increased by 4.9%. Furthermore, at the 32 nm technology node, a GNRFET-based structure was designed and compared to conventional CMOS and pure CNTFET-based IFFC-2SOA. It was discovered that the DC gain, phase margin, and power dissipation obtained by the CNTFET-based structure were superior to both. The DC Gain of CNTFET-IFFC-2SOA was 156.79% more than that of GNRFET-IFFC-2SOA. To confirm the workability of the proposed circuits, an integrator was designed and simulated as its application. The CNTFET-based IFFC-2SOA integrators showed better integration action than the traditional CMOS-IFFC-2SOA. [ABSTRACT FROM AUTHOR]

RNA polymerase II (RNAPII) transcription initiates bidirectionally at many human protein-coding genes. Sense transcription usually dominates and leads to messenger RNA production, whereas antisense transcription rapidly terminates. The basis for this directionality is not fully understood. Here, we show that sense transcriptional initiation is more efficient than in the antisense direction, which establishes initial promoter directionality. After transcription begins, the opposing functions of the endonucleolytic subunit of Integrator, INTS11, and cyclin-dependent kinase 9 (CDK9) maintain directionality. Specifically, INTS11 terminates antisense transcription, whereas sense transcription is protected from INTS11-dependent attenuation by CDK9 activity. Strikingly, INTS11 attenuates transcription in both directions upon CDK9 inhibition, and the engineered recruitment of CDK9 desensitises transcription to INTS11. Therefore, the preferential initiation of sense transcription and the opposing activities of CDK9 and INTS11 explain mammalian promoter directionality.

A second-order delta-sigma modulator (DSM) is proposed for readout integrated circuits of sensor applications requiring a small area and low-power consumption. The proposed second-order CIFF DSM with the architecture of cascaded-of-integrator feedforward (CIFF) basically consists of two integrators, a 3-bit quantizer, data-weighted averaging (DWA) circuit, and clock generator. The use of the 3-bit quantizer instead of the single-bit quantizer reduces the size of the feedback capacitor in the first integrator. The 3-bit quantizer is designed based on a successive approximation register analog-to-digital converter for small area and low power implementation. Furthermore, the proposed second-order CIFF DSM has a single supply without an additional reference driver while having a wide analog input voltage range with rail to rail. The proposed second-order CIFF DSM, implemented using a 130 nm 1-poly 6-metal CMOS process with a supply of 1.5 V, has an area of 0.096 mm2. It has a sampling frequency of 500 kHz for the implementation of an input bandwidth of 2 kHz and an oversampling ratio of 125. The measured peak signal-to-noise and distortion ratio is approximately 90 dB when the differential analog input signal has a frequency of 353 Hz and an amplitude of 1.2 Vpp. The measured dynamic range is approximately 96.3 dB. [ABSTRACT FROM AUTHOR]

The performance of free space optical communication system using multi-input multi-output orthogonal frequency division multiplexing (MIMO-OFDM) with a modified receiver structure is evaluated in this paper referred to as MIMO-OFDM-FSO with the modified receiver (MIMO-OFDM-FSO-MR). Utilizing equal gain combining the analysis is done by taking closed-form expression of average bit error rate and throughput. The simulation is performed to estimate various performance parameters such as throughput and bit error rate (BER) for the proposed MIMO-OFDM-FSO-MR model under different turbulence and weather conditions. Simulation reveals that the proposed model exhibits significant BER performance and provides better throughput and spectral efficiency as compared with conventional FSO systems, OFDM-FSO systems, MIMO-FSO systems and Low-density Parity Check (LDPC) coded MIMO-OFDM-FSO systems. [ABSTRACT FROM AUTHOR]

In this paper, linear quadratic regulator (LQR) control is applied to a Cuk converter, and mathematical modeling of the converter is done using state space averaging (SSA) in continuous conduction mode (CCM). The primary focus is to design a controller for the converter and maintain output voltage within 1% of the prescribed value when there is a unit step disturbance in the input. The design done by using MATLAB improved control of the converter from that with a conventional controller when tested with disturbances. The controller did not influence the system's stability, when assessed from the Bode plot. Despite the existence of disturbances, the findings prove the efficacy of the suggested approach, and the proposed controller can track the desired output voltage in less than 14 ms. The LQR control was developed for various applications of the Cuk converter, such as in a photovoltaic system and a wind system.

Current CMOS (Complementary metal–oxide–semiconductor) technology is no longer constrained in scaling by short channel effects. The semiconductor industry has developed a number of substitute technologies, including quantum-dot cellular automata, to get around these restrictions (QCA). In this study, a novel technique for developing digital differentiators and integrators is presented, employing QCA Technology as a key component. In order to design the digital differentiator focus has been given on no recursive simple tapped delay line differentiator called first difference differentiator and central difference differentiator. Further work has been done on design side of digital integrator. For this, time domain rectangular rule integrator has been realized with the support of QCA. By implementing the QCA, area of integrator circuit is obtained as 0.16 µm2 and 0.14 µm2 for 2 Bit and 4 Bit integrator circuit, respectively. Area occupied by central Difference differentiator is 0.52 µm2, 4 Bit F-D Differentiator occupied 0.53 µm2. Power calculation shows that power consumption is less than 6 nW in QCA-based integrator in comparison with CMOS technology. Further, it is observed that QCA-based digital differentiator and integrator have much reduced area compare to CMOS-based differentiator and integrator. These simple circuits can be further used in DSP filters like Cascaded Integrator-Comb (CIC). [ABSTRACT FROM AUTHOR]

The elongation stage of transcription by RNA polymerase II (RNA Pol II) is central to the regulation of gene expression in response to developmental and environmental cues in metazoan. Dysregulated transcriptional elongation has been associated with developmental defects as well as disease and aging processes. Decades of genetic and biochemical studies have painstakingly identified and characterized an ensemble of factors that regulate RNA Pol II elongation. This review summarizes recent findings taking advantage of genetic engineering techniques that probe functions of elongation factors in vivo. We propose a revised model of elongation control in this accelerating field by reconciling contradictory results from the earlier biochemical evidence and the recent in vivo studies. We discuss how elongation factors regulate promoter-proximal RNA Pol II pause release, transcriptional elongation rate and processivity, RNA Pol II stability and RNA processing, and how perturbation of these processes is associated with developmental disorders, neurodegenerative disease, cancer, and aging. Transcriptional elongation by RNA polymerase II is a crucial regulatory step in gene expression, and its dysregulation is linked to the mechanisms underlying human disease and aging processes. This review provides a comprehensive overview of both the current mechanistic understanding and the known biological relevance of RNA Pol II elongation factors. [ABSTRACT FROM AUTHOR]

In this paper, linear quadratic regulator (LQR) control is applied to a Cuk converter, and mathematical modeling of the converter is done using state space averaging (SSA) in continuous conduction mode (CCM). The primary focus is to design a controller for the converter and maintain output voltage within 1% of the prescribed value when there is a unit step disturbance in the input. The design done by using MATLAB improved control of the converter from that with a conventional controller when tested with disturbances. The controller did not influence the system's stability, when assessed from the Bode plot. Despite the existence of disturbances, the findings prove the efficacy of the suggested approach, and the proposed controller can track the desired output voltage in less than 14 ms. The LQR control was developed for various applications of the Cuk converter, such as in a photovoltaic system and a wind system. [ABSTRACT FROM AUTHOR]

In this work, we propose, design and simulate a new configuration of Class AB dual output (DO) second‐generation current conveyor (DO‐CCII). Two versions of this new DO‐CCII configuration are proposed: one employing 32 nm technology‐based carbon nanotube field effect transistor (CNTFET) and the other employing the conventional complementary MOS (CMOS). The HSPICE simulation studies have been performed to evaluate key performance parameters like current and voltage gains, voltage and current bandwidths (BW), the port X and Y resistances, total harmonic distortion of both proposed DO‐CCII configurations. It has been seen that ~388× and ~215× improvement in current and voltage BWs respectively have been achieved in the proposed CNTFET based DO‐CCII (CNTFET‐DO‐CCII) configuration in comparison to its CMOS counterpart (CMOS‐DO‐CCII). Similarly, desired port X and Y resistances have resulted substantially in the CNTFET based DO‐CCII configuration. Further, to evaluate the performance of proposed DO‐CCIIs, some applications such as sinusoidal oscillator, integrator and differentiator have been simulated and compared. In the designed oscillators, the effect of temperature on oscillating frequency has also been studied. A temperature insensitive behavior has been seen in the CNTFET‐DO‐CCII for a wide frequency range. Furthermore, it has been found that the optimization of number of CNTs, CNT diameter and inter‐CNT pitch of CNTFETs improve the performance of the proposed DO‐CCII. [ABSTRACT FROM AUTHOR]

The judicious utilization of available daylight for indoor illumination is an indispensable requirement for buildings. A daylight pipe is considered an appropriate daylighting system to bring daylight in a deep-plan indoor space. Mathematical models are available to predict horizontal illuminance contributed by daylight pipe as a function of global horizontal illuminance. Since light collection and propagation characteristics of direct and diffuse light through a highly specular light guiding system are different, direct and diffuse components of global illuminance are to be considered separately. This paper presents a mathematical formulation for estimation of the delivered instantaneous lumen by a straight tubular hollow daylight pipe system as a function of instantaneous global horizontal illuminance considering direct and diffuse components separately which is applicable for mostly clear sky conditions prevail in a tropical region. Experimental validation of the proposed formulation is conducted under the real sky and the root means square deviation and the percentage deviation of the predicted lumen from the measured lumen are found at 23.83 lumens and 1.17%, respectively, while the correlation coefficient is obtained as 0.9967. Finally, the application of the proposed methodology is demonstrated with DLPs of different configurations for four representative days using measured solar radiation data under Indian sky conditions. [ABSTRACT FROM AUTHOR]

Metazoans guard their germlines against transposons and other foreign transcripts with PIWI-interacting RNAs (piRNAs). Due to the robust heritability of the silencing initiated by piRNAs in Caenorhabditis elegans (C. elegans), previous screens using C. elegans were strongly biased to uncover members of this pathway in the maintenance process but not in the initiation process. To identify novel piRNA pathway members, we have utilized a sensitized reporter strain which detects defects in initiation, amplification, or regulation of piRNA silencing. Using our reporter, we have identified Integrator complex subunits, nuclear pore components, protein import components, and pre-mRNA splicing factors as essential for piRNA-mediated gene silencing. We found the small nuclear processing cellular machine termed the Integrator complex is required for both type I and type II piRNA production. Notably, we identified a role for nuclear pore and nucleolar components NPP-1/Nup54, NPP-6/Nup160, NPP-7/Nup153, and FIB-1 in promoting the perinuclear localization of anti-silencing CSR-1 Argonaute, as well as a role for Importin factor IMA-3 in nuclear localization of silencing Argonaute HRDE-1. Together, we have shown that piRNA silencing in C. elegans is dependent on evolutionarily ancient RNA processing machinery that has been co-opted to function in the piRNA-mediated genome surveillance pathway. [ABSTRACT FROM AUTHOR]

In this paper, an active implementation of a differential voltage current conveyor (DVCC) based on a low-pass filter operating in the fractional order domain is presented. The transfer function for a fractional order system is dependent on the rational approximation of sα. Different methods used for calculating the rational approximation, including Carlson, Elkhazalil, and curve fitting, are evaluated here. Finally, to validate the theoretical results, a fractional order Butterworth filter is simulated in the Pspice environment using the 0.5 micrometer CMOS technology with an R-C network-based fractional order capacitor. Additionally, using the Monte Carlo analysis, the impact of current and voltage faults on DVCC response is investigated. It has been inferred that realization with a wider bandwidth is possible. [ABSTRACT FROM AUTHOR]

In this paper, an active implementation of a differential voltage current conveyor (DVCC) based on a low-pass filter operating in the fractional order domain is presented. The transfer function for a fractional order system is dependent on the rational approximation of sα. Different methods used for calculating the rational approximation, including Carlson, Elkhazalil, and curve fitting, are evaluated here. Finally, to validate the theoretical results, a fractional order Butterworth filter is simulated in the Pspice environment using the 0.5 micrometer CMOS technology with an R-C network-based fractional order capacitor. Additionally, using the Monte Carlo analysis, the impact of current and voltage faults on DVCC response is investigated. It has been inferred that realization with a wider bandwidth is possible.

In this study, we considered the regulator problem for discrete‐valued input control with an integrator. In some cases, the state does not converge to the origin in the discrete‐valued input control when the plant possesses an integrator. Therefore, a method that adds minor feedback control to the integrated value of the input was proposed. However, the effect of the feedback control of the integrated value of the input was not theoretically interpreted, and the feedback gain for the state of the integrated value of the input was designed with trial and error. Herein, we clarify the effect of feedback control on the integrated value of the input. Furthermore, we reveal that the trade‐off between the steady‐state and transient performances is controlled by one parameter, that is, the feedback gain for the integrated value of the input. [ABSTRACT FROM AUTHOR]

Nowadays autonomous measurement systems are applied for the determination of the cinematic parameters of object motion. The principle of operation of such systems is based on the measurement of the object motion acceleration with the further integration of the accelerometer's signal. These measurement systems are called inertial systems. Platform-less inertial systems are widely used for these purposes. Their specific property is an installation of the primary information sensors directly on the constructive elements of the object case. The inertial measurement systems have also a negative feature to accumulate constantly the errors of the system. [ABSTRACT FROM AUTHOR]

Integrator is a metazoan-specific protein complex capable of inducing termination at all RNAPII-transcribed loci. Integrator recognizes paused, promoter-proximal RNAPII and drives premature termination using dual enzymatic activities: an endonuclease that cleaves nascent RNA and a protein phosphatase that removes stimulatory phosphorylation associated with RNAPII pause release and productive elongation. Recent breakthroughs in structural biology have revealed the overall architecture of Integrator and provided insights into how multiple Integrator modules are coordinated to elicit termination effectively. Furthermore, functional genomics and biochemical studies have unraveled how Integrator-mediated termination impacts protein-coding and noncoding loci. Here, we review the current knowledge about the assembly and activity of Integrator and describe the role of Integrator in gene regulation, highlighting the importance of this complex for human health. Integrator broadly drives promoter-proximal termination of RNAPII through phosphatase and RNA endonuclease activities. Here, we summarize recent work illustrating the molecular underpinnings of Integrator activity. Furthermore, we describe the critical role Integrator plays in human health and disease through its regulation of gene expression and noncoding RNA synthesis. [ABSTRACT FROM AUTHOR]

Transformer excitation magnetizing curves (TEMC) reflect the dynamic operation characteristics of iron-core materials. Using numerical analysis and the waveform recording function of digital oscilloscopes, we developed a cost-effective method for determining the TEMC. This approach eliminates the need for conventional analog integrator circuits. To address the potential obstacles to the digital generation of TEMC—namely, curve offsets and curve transients—we proposed solutions involving Fourier filtering and determining the initial point of integration. The results indicate that the proposed approach yields results consistent with those of conventional analog integrator circuits and highlight its promise for applications in data processing. [ABSTRACT FROM AUTHOR]

First paragraphs: Why do industrial agricultural operations continue to displace smaller family farms in spite of their continued pollution of the natural environment and degradation of rural communi­ties? Large-scale, specialized agricultural operations, such as concentrated animal feeding operations (or CAFOs), persist because they have an economic advantage over smaller, diversified farming opera­tions. They have higher ecological and social costs but lower economic costs. This economic advan­tage is commonly referred to as economies of scale. In economic theory, there are two types of economies of scale. Internal economies of scale refer to differences in the costs of production associated with different sizes of production units. In animal agriculture, “scale” refers to the number of hogs, poultry, milk cows, or beef cattle in a single farming operation or production unit. In field crop and pasture-based animal production, scale refers to the acres of land in a single production unit. External economies of scale, on the other hand, refer to differences such as the costs of fertilizer or feed, or the cost of complying with government regulations, for different sizes of management units. Management units may include one or more production units under single man­agement or control (Ross, 2022). A single farm or production unit may comprise multiple parcels of land, but a farm management unit may comprise multiple farms that are managed as a single economic entity or unit. . . .

Operational amplifiers (op-amps) are generally used for actualizing simple and complex electronic circuits in the subject of analogue electronics. In an effort to improve the teaching of op-amps in electronics engineering curricula, op-amp circuits in various configurations are often used for experiments in laboratory sessions so that students can acquire certain psychomotor and cognitive skills by constructing circuit connections and analyzing input–output waveforms. As a result, multiple configurations of operational amplifier circuits are often needed, requiring multiple sets of experimental boards or circuits for each experiment. This is usually not cost effective, requires more consumable electronic components, requires more maintenance and storage space in facilities, and is less user friendly for the students. Therefore, the aim of this research is to design a single, compact, and easy-to-replicate experimental board that can be converted into multiple configurations of the LM741 operational amplifier, comprising an inverting amplifier, a noninverting amplifier, a voltage follower, a summing amplifier, a differential amplifier, a differentiator, and an integrator, with minimal electronic components at a cost lower than EUR 10. The experimental board was tested with a constant input voltage of 1.0 V AC and a switching frequency of 1.0 kHz. It is capable of producing an output voltage corresponding to the individual operational amplifier configurations and can thus be used as a facilitating module for teaching and learning activities in the field of analogue electronics. [ABSTRACT FROM AUTHOR]

Summary: In this paper, a stator current‐based model reference adaptive system (SCMRAS) for indirect vector control of induction motor fed to hybrid electric vehicle (HEV) for improving the transient response during limp home period has been proposed. In the proposed SCMRAS, the measured stator currents are employed in voltage model to eliminate the integrator in reference model. The stator currents are estimated and are compared with actual current components to estimate the rotor speed. Further, to improve the performance of SCMRAS during limp home period, the PI controller in the adaptation mechanism is replaced with type 2 fuzzy logic controller (T2FLC). The prototype model of the proposed SCMRAS using dSPACE DS 1104 R&D controller board has been developed for implementing speed sensorless indirect vector control of induction motor drive. The performance of SCMRAS and proposed SCMRAS using T2FLC estimators during limp home period is compared. [ABSTRACT FROM AUTHOR]

In many contexts of scientific computing and engineering science, phenomena are monitored over time and data are collected as time-series. Plenty of algorithms have been proposed in the field of time-series data mining, many of them based on deep learning techniques. High-fidelity simulations of complex scenarios are truly computationally expensive and a real-time monitoring and control could be efficiently achieved by the use of artificial intelligence. In this work we build accurate data-driven models of a two-phase transient flow in a heated channel, as usually encountered in heat exchangers. The proposed methods combine several artificial neural networks architectures, involving standard and transposed deep convolutions. In particular, a very accurate real-time integrator of the system has been developed. [ABSTRACT FROM AUTHOR]

The best-studied mechanism of eukaryotic RNA polymerase II (RNAPII) transcriptional termination involves polyadenylation site-directed cleavage of the nascent RNA. The RNAPII-associated cleavage product is then degraded by XRN2, dislodging RNAPII from the DNA template. In contrast, prokaryotic RNAP and eukaryotic RNAPIII often terminate directly at T-tracts in the coding DNA strand. Here, we demonstrate a similar and omnipresent capability for mammalian RNAPII. Importantly, this termination mechanism does not require upstream RNA cleavage. Accordingly, T-tract-dependent termination can take place when XRN2 cannot be engaged. We show that T-tracts can terminate snRNA transcription independently of RNA cleavage by the Integrator complex. Importantly, we found genome-wide termination at T-tracts in promoter-proximal regions but not within protein-coding gene bodies. XRN2-dependent termination dominates downstream from protein-coding genes, but the T-tract process is sometimes used. Overall, we demonstrate global DNA-directed attrition of RNAPII transcription, suggesting that RNAPs retain the potential to terminate over T-rich sequences throughout evolution., (© 2024 Davidson et al.; Published by Cold Spring Harbor Laboratory Press.)

The biological purpose of Integrator and RNA polymerase II (RNAPII) promoter-proximal pausing remains uncertain. Here, we show that loss of INTS6 in human cells results in increased interaction of RNAPII with proteins that can mediate its dissociation from the DNA template, including the CRL3 ARMC5 E3 ligase, which ubiquitylates CTD serine 5 -phosphorylated RPB1 for degradation. ARMC5-dependent RNAPII ubiquitylation is activated by defects in factors acting at the promoter-proximal pause, including Integrator, DSIF, and capping enzyme. This ARMC5 checkpoint normally curtails a sizeable fraction of RNAPII transcription, and ARMC5 knockout cells produce more uncapped transcripts. When both the Integrator and CRL3 ARMC5 turnover mechanisms are compromised, cell growth ceases and RNAPII with high pausing propensity disperses from the promoter-proximal pause site into the gene body. These data support a model in which CRL3 ARMC5 functions alongside Integrator in a checkpoint mechanism that removes faulty RNAPII complexes at promoter-proximal pause sites to safeguard transcription integrity., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Abstract Background Notch signaling drives many aspects of neoplastic phenotype. Here, we report that the Integrator complex (INT) is a new component of the Notch transcriptional supercomplex. Together with Notch Activation Complex Kinase (NACK), INT activates Notch1 target genes by driving RNA polymerase II (RNAPII)-dependent transcription, leading to tumorigenesis. Methods Size exclusion chromatography and CBF-1/RBPJ/Suppressor of Hairless/Lag-1 (CSL)-DNA affinity fast protein liquid chromatography (FPLC) was used to purify Notch/CSL-dependent complexes for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Chromatin immunoprecipitation (ChIP) and quantitative polymerase chain reaction (qPCR) were performed to investigate transcriptional regulation of Notch target genes. Transfection of Notch Ternary Complex components into HEK293T cells was used as a recapitulation assay to study Notch-mediated transcriptional mechanisms. Gene knockdown was achieved via RNA interference and the effects of protein depletion on esophageal adenocarcinoma (EAC) proliferation were determined via a colony formation assay and murine xenografts. Western blotting was used to examine expression of INT subunits in EAC cells and evaluate apoptotic proteins upon INT subunit 11 knockdown (INTS11 KD). Gene KD effects were further explored via flow cytometry. Results We identified the INT complex as part of the Notch transcriptional supercomplex. INT, together with NACK, activates Notch-mediated transcription. While NACK is required for the recruitment of RNAPII to a Notch-dependent promoter, the INT complex is essential for RNAPII phosphorylated at serine 5 (RNAPII-S5P), leading to transcriptional activation. Furthermore, INT subunits are overexpressed in EAC cells and INTS11 KD results in G2/M cell cycle arrest, apoptosis, and cell growth arrest in EAC. Conclusions This study identifies the INT complex as a novel co-factor in Notch-mediated transcription that together with NACK activates Notch target genes and leads to cancer cell proliferation. Video abstract

INTS11 and CPSF73 are metal-dependent endonucleases for Integrator and pre-mRNA 3′-end processing, respectively. Here, we show that the INTS11 binding partner BRAT1/CG7044, a factor important for neuronal fitness, stabilizes INTS11 in the cytoplasm and is required for Integrator function in the nucleus. Loss of BRAT1 in neural organoids leads to transcriptomic disruption and precocious expression of neurogenesis-driving transcription factors. The structures of the human INTS9-INTS11-BRAT1 and Drosophila dIntS11-CG7044 complexes reveal that the conserved C terminus of BRAT1/CG7044 is captured in the active site of INTS11, with a cysteine residue directly coordinating the metal ions. Inspired by these observations, we find that UBE3D is a binding partner for CPSF73, and UBE3D likely also uses a conserved cysteine residue to directly coordinate the active site metal ions. Our studies have revealed binding partners for INTS11 and CPSF73 that behave like cytoplasmic chaperones with a conserved impact on the nuclear functions of these enzymes. [Display omitted] • BRAT1/CG7044 is a binding partner that stabilizes INTS11 in the cytoplasm • BRAT1/CG7044 is required for Integrator function in the nucleus • The conserved C terminus of BRAT1-CG7044 is captured in the INTS11 active site • UBE3D is a binding partner of CPSF73 with a similar mechanism of action Lin et al. reveal BRAT1/CG7044 as a binding partner for the INTS11 endonuclease that behaves like a cytoplasmic chaperone and is required for Integrator function in the nucleus. UBE3D is also identified as a binding partner for the CPSF73 endonuclease in pre-mRNA 3′-end processing, with a similar mechanism of action. [ABSTRACT FROM AUTHOR]

The Rogowski coil transducer's integrator will amplify the transmission error, which may lead to mal-operation of the relay. In this paper, a novel method of line relay based on the differential output of Rogowski coil is proposed. Besides, an improved fast distance relay based on the differential output of Rogowski coil is presented. Based on the equal transfer process of transmission lines(ETPTL), the voltage signal should be processed by the virtual Rogowski coil to solve the inconsistent transfer problem between voltage and current. Furthermore, this paper also presents the reconstruction method of voltage at the fault point and the least-square algorithm to solve the R-L differential equation, so as to improve the algorithm accuracy. Furthermore, the equal transfer process error of the virtual Rogowski coil is analyzed in the paper, and the design principle of low pass filter is also proposed. Simulation results and experiment results demonstrate that the improved fast distance relay scheme has high operation speed, which is signed prior to other distance relay schemes. The proposed method also provides a valuable reference for other types of transmission line relay based on Rogowski coil transducer. [ABSTRACT FROM AUTHOR]