Intacs

The metazoan-specific Integrator complex catalyzes 3' end processing of small nuclear RNAs (snRNAs) and premature termination that attenuates the transcription of many protein-coding genes. Integrator has RNA endonuclease and protein phosphatase activities, but it remains unclear if both are required for complex function. Here, we show IntS6 (Integrator subunit 6) over-expression blocks Integrator function at a subset of Drosophila protein-coding genes, although having no effect on snRNAs or attenuation of other loci...

PURPOSE: The aim of this study was to analyze changes in corneal biomechanical properties after implantation of intracorneal ring segments (ICRSs) in keratectasia. METHODS: This retrospective single-center study included 112 patient eyes that underwent femtosecond laser-assisted ICRS implantation (Intacs SK; Addition Technology Inc, Des Plaines, IL) for keratectasia. Biomechanical analysis was performed using the Ocular Response Analyzer (ORA; Reichert Inc, Depew, NY), with determination of corneal resistance factor, corneal hysteresis, and Keratoconus Match Index, as well as by Corvis ST (OCULUS, Wetzlar, Germany), with determination of stiffness parameter A1, Ambrosio relational thickness to the horizontal profile (Arth), integrated radius, deformation amplitude ratio, and stress-strain index as well as Corvis Biomechanical Index and Tomographic Biomechanical Index...

A recent Nature paper by Xu et al.1 describes an important link between RNA polymerase II promoter-proximal pausing and genome stability orchestrated by liquid droplet formation to reduce unwanted R-loop accumulation.

While several inhibitors targeting RNA polymerase II (Pol II) kinases have been applied for inhibiting RNA Pol II phosphorylation, there are few approaches for inducing RNA Pol II hyperphosphorylation. Here, we present a protocol for constructing the INTS8 degradation tag (dTAG) system combined with ectopic expression of N-terminally truncated INTS8 (INTS8-ΔN) in DLD-1 cells. We describe steps for INTS8-dTAG cell line construction, validation of knockin and degradation, and INTS8-ΔN rescue. We then detail validation of RNA Pol II phosphorylation upregulation...

The stable association between SOSS and INTAC maintains genome stability by limiting R-loop accumulation.

The proper regulation of transcription is essential for maintaining genome integrity and executing other downstream cellular functions1,2 . Here we identify a stable association between the genome-stability regulator sensor of single-stranded DNA (SOSS)3 and the transcription regulator Integrator-PP2A (INTAC)4-6 . Through SSB1-mediated recognition of single-stranded DNA, SOSS-INTAC stimulates promoter-proximal termination of transcription and attenuates R-loops associated with paused RNA polymerase II to prevent R-loop-induced genome instability...

Cyclin-dependent kinase 12 (CDK12) interacts with cyclin K to form a functional nuclear kinase that promotes processive transcription elongation through phosphorylation of the C-terminal domain of RNA polymerase II (Pol II). To gain a comprehensive understanding of CDK12's cellular function, we used chemical genetic and phosphoproteomic screening to identify a landscape of nuclear human CDK12 substrates, including regulators of transcription, chromatin organization, and RNA splicing. We further validated LEO1, a subunit of the polymerase-associated factor 1 complex (PAF1C), as a bona fide cellular substrate of CDK12...

Gene expression in metazoans is controlled by promoter-proximal pausing of RNA polymerase II, which can undergo productive elongation or promoter-proximal termination. Integrator-PP2A (INTAC) plays a crucial role in determining the fate of paused polymerases, but the underlying mechanisms remain unclear. Here, we establish a rapid degradation system to dissect the functions of INTAC RNA endonuclease and phosphatase modules. We find that both catalytic modules function at most if not all active promoters and enhancers, yet differentially affect polymerase fate...

No abstract text is available yet for this article.

PURPOSE: To compare the clinical outcomes of intracorneal ring segment (ICRS) implantation in eyes with advanced vs. mild/moderate keratoconus (KCN). METHODS: A retrospective analysis of 141 eyes of 111 patients with KCN who underwent ICRS implantation. Preoperative maximum keratometry (Kmax) was <57 diopters (D) in 70 eyes and >57 D in 71 eyes. Postoperatively, corrected distance visual acuity (CDVA), Kmax, and intraoperative and postoperative complications were assessed at 1 day, 1 month, and 1 year...

Intrinsic protein flexibility is of overwhelming relevance for intermolecular recognition and adaptability of highly dynamic ensemble of complexes, and the phenomenon is essential for the understanding of numerous biological processes. These conformational ensembles-encounter complexes-lack a unique organization, which prevents the determination of well-defined high resolution structures. This is the case for complexes involving the oncoprotein SET/template-activating factor-Iβ (SET/TAF-Iβ), a histone chaperone whose functions and interactions are significantly affected by its intrinsic structural plasticity...

Master transcription factors (TFs) directly regulate present and future cell states by binding DNA regulatory elements and driving gene-expression programs. Their abundance influences epigenetic priming to different cell fates at the chromatin level, especially in the context of differentiation. In order to link TF protein abundance to changes in TF motif accessibility and open chromatin, we developed InTAC-seq, a method for simultaneous quantification of genome-wide chromatin accessibility and intracellular protein abundance in fixed cells...

PURPOSE: To evaluate management of keratoconic eyes with anterior stromal necrosis overlying the intracorneal ring segment (ICRS), by either ICRS explantation alone or exchange with corneal allogenic intrastromal ring segments (CAIRS). METHODS: Among 643 Intacs SK (Additional Technology, Inc) inserted at one institutional center, 16 eyes (15 patients) with overlying spontaneous anterior stromal necrosis were identified. Data included size of stromal defect and refractive and topographical findings before ICRS insertion, before anterior stromal necrosis, and 6 months after intervention...

PURPOSE: To determine the effect of a single-segment intrastromal corneal ring segment (ICRS; Intacs SK) on early keratoconus (KCN) and pellucid marginal degeneration (PMD). METHODS: It is a prospective interventional study. One hundred twenty-four eyes (99 patients) with KCN and 36 eyes (26 patients) with PMD at early stage (the maximum keratometric reading less than 55 diopters) were included to ICRS implantation using femtosecond laser at a tertiary university-based hospital and a private outpatient center...

Pleiotropic transcription regulator RNA polymerase II (Pol II)-associated factor 1 (PAF1) governs multiple transcriptional steps and the deposition of several epigenetic marks. However, it remains unclear how ultimate transcriptional outcome is determined by PAF1 and whether it relates to PAF1-controlled epigenetic marks. We use rapid degradation systems and reveal direct PAF1 functions in governing pausing partially by recruiting Integrator-PP2A (INTAC), in addition to ensuring elongation. Following acute PAF1 degradation, most destabilized polymerase undergoes effective release, which presumably relies on skewed balance between INTAC and P-TEFb, resulting in hyperphosphorylated substrates including SPT5...

Objective: An animal model of collagen-induced arthritis (CIA) was used to investigate the effects of norepinephrine (NE) and α1-adrenoreceptor (α1-AR) on Treg cells in CIA. Methods: Thirty-two male DBA/1 mice were randomly divided into control group and CIA model group. CIA was prepared by intradermal injection of collagen type II (CII, 100 μl) at the tail base of DBA/1 mice. On the 41th day following primary immunization, co-expression of CD4+ T and α1-AR in mouse spleens was observed. Protein expressions of α1-AR in the ankle joints and the spleens of mice were measured by Western blot analysis...

Intrastromal corneal ring segments (ICRSs) are an effective treatment for stabilizing and normalizing corneal shape in patients with keratoconus and other corneal ectasias. Intraoperative segment perforation through the corneal endothelium into the anterior chamber (AC) is an uncommon but known complication. However, perforation into the AC postoperatively is an exceedingly rare complication with only 3 reported cases in the literature. One case was due to Descemet membrane detachment and another due to ocular trauma...

Transcription progression is governed by multitasking regulators including SPT5, an evolutionarily conserved factor implicated in virtually all transcriptional steps from enhancer activation to termination. Here we utilize a rapid degradation system and reveal crucial functions of SPT5 in maintaining cellular and chromatin RNA polymerase II (Pol II) levels. Rapid SPT5 depletion causes a pronounced reduction of paused Pol II at promoters and enhancers, distinct from negative elongation factor (NELF) degradation resulting in short-distance paused Pol II redistribution...

PURPOSE: To describe a long-term outcome of a new technique using the femtosecond laser to assist with keratopigmentation (corneal tattooing). METHOD: Nineteen eyes of 18 patients with a history of significant visual disability related to iris defects underwent femtosecond laser-assisted keratopigmentation (FAK) for functional restoration of their visual disabilities. Postoperative visual acuity and symptomatic resolution were collected during their postoperative visits...

SIGNIFICANCE: Keratoconus can manifest asymmetrically, affecting binocularity and becoming a refractive problem that is sometimes complex to solve. We propose a therapeutic approach for correction of keratoconus based on parallel implantation of a second intrastromal corneal ring segment (ICRS). PURPOSE: This study aimed to improve the refractive status of a patient affected with advanced bilateral keratoconus using implantation of a second ICRS and a phakic intraocular lens...