Actin binding to WH2 domains regulates nuclear import of the multifunctional actin regulator JMY

© The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Molecular Biology of the Cell 23 (2012): 853-863, doi:10.1091/mbc.E11-12-0992.Junction-mediating and regulatory protein (JMY) is a regulator of both transcription and actin filament assembly. In response to DNA damage, JMY accumulates in the nucleus and promotes p53-dependent apoptosis. JMY's actin-regulatory activity relies on a cluster of three actin-binding Wiskott–Aldrich syndrome protein homology 2 (WH2) domains that nucleate filaments directly and also promote nucleation activity of the Arp2/3 complex. In addition to these activities, we find that the WH2 cluster overlaps an atypical, bipartite nuclear localization sequence (NLS) and controls JMY's subcellular localization. Actin monomers bound to the WH2 domains block binding of importins to the NLS and prevent nuclear import of JMY. Mutations that impair actin binding, or cellular perturbations that induce actin filament assembly and decrease the concentration of monomeric actin in the cytoplasm, cause JMY to accumulate in the nucleus. DNA damage induces both cytoplasmic actin polymerization and nuclear import of JMY, and we find that damage-induced nuclear localization of JMY requires both the WH2/NLS region and importin β. On the basis of our results, we propose that actin assembly regulates nuclear import of JMY in response to DNA damage.This work was supported by grants from the National Institutes of Health, an American Heart Association Predoctoral Fellowship (J.B.Z.), the Robert Day Allen Fellowship Fund (J.B.Z.), and a National Science Foundation Predoctoral Fellowship (B.B.)

NR5A1 gene variants repress the ovarian-specific WNT signaling pathway in 46,XX disorders of sex development patients

Several recent reports have described a missense variant in the gene NR5A1 (c.274C>T; p.Arg92Trp) in a significant number of 46,XX ovotesticular or testicular disorders of sex development (DSDs) cases. The affected residue falls within th

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

Effects of Instrument Assisted Soft-Tissue Mobilization on Neuromuscular Control for Chronic Ankle Instability

Ankle injuries can cause Chronic Ankle Instability (CAI), causing deficits in ankle mobility and stability. The fibularis longus can have decreased neural excitability associated with CAI. Instrument assisted soft-tissue mobilization (IASTM) is a therapy technique that assists with scar tissue, fascia mobilization, and tissue healing. The objective of our study was to examine the effects of the application of IASTM to the fibularis longus on neuromuscular control (NMC) in individuals with CAI. The independent variables were session (application of IASTM, control) and time (pre- and post-test). The dependent variable was the Y-balance test (YBT) reach distances. We determined if participants had CAI using the ankle instability instrument. IASTM was applied to the fibularis longus muscle for 90 seconds during the intervention, and participants sat for 2 minutes during the control. Pre-test and post-test measurements were taken using the YBT with the best score of 3 trials used for analysis. The interaction between session and time for anterior reach was significant (F1,14=5.256, P=.038, η2=.273). The interaction between session and time was not significant for posteromedial (F1,14=.251, P=.624, η2=.018, 1-β=.075) or posterolateral (F1,14=1.166, P=.298, η2=.077, 1-β=.172) reaches. Our findings suggested that IASTM to the fibularis longus does not immediately improve all aspects of dynamic NMC. The 3.5 cm increase in anterior reach could decrease the risk of non-contact injury if there is asymmetry between injured and uninjured limbs. Keywords: Ankle sprains, Y-balance test, fibularis longus, postural control, manual therap

Effects of Instrument Assisted Soft-Tissue Mobilization on Dynamic Balance in Those with Chronic Ankle Instability

Our objective was to examine the effectiveness of IASTM application to the FL on dynamic balance in individuals with CAI. Fifteen individuals (seven females, eight males, age = 26.07 ± 9.18 years, mass = 87.33 ± 24.07 kg, height = 178.83 ± 12.83 cm) with CAI, as determined by the Ankle Instability Instrument (AII) volunteered to participate. Participants completed two counterbalanced sessions (experimental and control), and we recorded measurements at two time points (pre- and post-). The application of IASTM to the FL muscle was carried out using Técnica Gavilán® instruments for 90 s during the intervention, and participants sat for 2 min during the control session. Dynamic balance was assessed using the Y-balance test (YBT). The interaction between session and time for anterior reach was significant (F1,14 = 5.26, p = 0.04, η2 = 0.27). Post-hoc tests revealed farther reach distances at post-test (71.02 ± 9.45 cm) compared to pre-test (66.57 ± 10.87 cm) when IASTM was applied (p = 0.02, Mean Difference = 4.45 cm, CI95 = 0.71–8.19 cm, Cohen’s d = 0.44). The interaction between session and time was not significant for posteromedial (F1,14 = 0.25, p = 0.62, η2 = 0.02, 1 − β = 0.08) or posterolateral reaches (F1,14 = 1.17, p = 0.30, η2 = 0.08, 1 − β = 0.17). The application of IASTM to the FL improved anterior reach of the YBT, but not posterolateral or posteromedial reaches in individuals with CAI. However, the 4.45 cm increase in anterior reach could have clinical implications for improved function

Effects of Instrument Assisted Soft-Tissue Mobilization on Dynamic Balance in Those with Chronic Ankle Instability

Our objective was to examine the effectiveness of IASTM application to the FL on dynamic balance in individuals with CAI. Fifteen individuals (seven females, eight males, age = 26.07 ± 9.18 years, mass = 87.33 ± 24.07 kg, height = 178.83 ± 12.83 cm) with CAI, as determined by the Ankle Instability Instrument (AII) volunteered to participate. Participants completed two counterbalanced sessions (experimental and control), and we recorded measurements at two time points (pre- and post-). The application of IASTM to the FL muscle was carried out using Técnica Gavilán® instruments for 90 s during the intervention, and participants sat for 2 min during the control session. Dynamic balance was assessed using the Y-balance test (YBT). The interaction between session and time for anterior reach was significant (F1,14 = 5.26, p = 0.04, η2 = 0.27). Post-hoc tests revealed farther reach distances at post-test (71.02 ± 9.45 cm) compared to pre-test (66.57 ± 10.87 cm) when IASTM was applied (p = 0.02, Mean Difference = 4.45 cm, CI95 = 0.71–8.19 cm, Cohen’s d = 0.44). The interaction between session and time was not significant for posteromedial (F1,14 = 0.25, p = 0.62, η2 = 0.02, 1 − β = 0.08) or posterolateral reaches (F1,14 = 1.17, p = 0.30, η2 = 0.08, 1 − β = 0.17). The application of IASTM to the FL improved anterior reach of the YBT, but not posterolateral or posteromedial reaches in individuals with CAI. However, the 4.45 cm increase in anterior reach could have clinical implications for improved function

A novel heterozygous variant in FGF9 associated with previously unreported features of multiple synostosis syndrome 3

Human multiple synostoses syndrome 3 is an autosomal dominant disorder caused by pathogenic variants in FGF9. Only two variants have been described in FGF9 in humans so far, and one in mice. Here we report a novel missense variant c.566C>G, p.(Pro189Arg) in FGF9. Functional studies showed this variant impairs FGF9 homodimerization, but not FGFR3c binding. We also review the findings of cases reported previously and report on additional features not described previously

Generation and mutational analysis of a transgenic mouse model of human SRY

SRY is the Y-chromosomal gene that determines male sex development in humans and most other mammals. After three decades of study, we still lack a detailed understanding of which domains of the SRY protein are required to engage the pathway of gene activity leading to testis development. Some insight has been gained from the study of genetic variations underlying differences/disorders of sex determination (DSD), but the lack of a system of experimentally generating SRY mutations and studying their consequences in vivo has limited progress in the field. To address this issue, we generated a mouse model carrying a human SRY transgene able to drive testis determination in XX mice. Using CRISPR-Cas9 gene editing, we generated novel genetic modifications in each of SRY's three domains (N-terminal, HMG box, and C-terminal) and performed a detailed analysis of their molecular and cellular effects on embryonic testis development. Our results provide new functional insights unique to human SRY and present a versatile and powerful system in which to functionally analyze variations of SRY including known and novel pathogenic variants found in DSD

Ovotesticular disorders of sex development in FGF9 mouse models of human synostosis syndromes

In mice, male sex determination depends on FGF9 signalling via FGFR2c in the bipotential gonads to maintain the expression of the key testis gene SOX9. In humans, however, while FGFR2 mutations have been linked to 46,XY disorders of sex development (DSD), the role of FGF9 is unresolved. The only reported pathogenic mutations in human FGF9, FGF9S99N and FGF9R62G, are dominant and result in craniosynostosis (fusion of cranial sutures) or multiple synostoses (fusion of limb joints). Whether these synostosis-causing FGF9 mutations impact upon gonadal development and DSD etiology has not been explored. We therefore examined embryonic gonads in the well-characterized Fgf9 missense mouse mutants, Fgf9S99N and Fgf9N143T, which phenocopy the skeletal defects of FGF9S99N and FGF9R62G variants, respectively. XY Fgf9S99N/S99N and XY Fgf9N143T/N143T fetal mouse gonads showed severely disorganized testis cords and partial XY sex reversal at 12.5\ua0days post coitum (dpc), suggesting loss of FGF9 function. By 15.5 dpc, testis development in both mutants had partly recovered. Mitotic analysis in vivo and in vitro suggested that the testicular phenotypes in these mutants arise in part through reduced proliferation of the gonadal supporting cells. These data raise the possibility that human FGF9 mutations causative for dominant skeletal conditions can also lead to loss of FGF9 function in the developing testis, at least in mice. Our data suggest that, in humans, testis development is largely tolerant of deleterious FGF9 mutations which lead to skeletal defects, thus offering an explanation as to why XY DSDs are rare in patients with pathogenic FGF9 variants

Mutant NR5A1/SF-1 in patients with disorders of sex development shows defective activation of the SOX9 TESCO enhancer

International audienceNuclear receptor subfamily 5 group A member 1/Steroidogenic factor 1 (NR5A1; SF-1; Ad4BP) mutations cause 46,XY disorders of sex development (DSD), with phenotypes ranging from developmentally mild (e.g., hypospadias) to severe (e.g., complete gonadal dysgenesis). The molecular mechanism underlying this spectrum is unclear. During sex determination, SF-1 regulates SOX9 (SRY [sex determining region Y]-box 9) expression. We hypothesized that SF-1 mutations in 46,XY DSD patients affect SOX9 expression via the Testis-specific Enhancer of Sox9 core element, TESCO. Our objective was to assess the ability of 20 SF-1 mutants found in 46,XY DSD patients to activate TESCO. Patient DNA was sequenced for SF-1 mutations and mutant SF-1 proteins were examined for transcriptional activity, protein expression, sub-cellular localization and in silico structural defects. Fifteen of the 20 mutants showed reduced SF-1 activation on TESCO, 11 with atypical sub-cellular localization. Fourteen SF-1 mutants were predicted in silico to alter DNA, ligand or cofactor interactions. Our study may implicate aberrant SF-1-mediated transcriptional regulation of SOX9 in 46,XY DSDs