Epidermal Growth Factor Receptor Dependence of Radiation-induced Transcription Factor Activation in Human Breast Carcinoma Cells

Ionizing radiation (1–5 Gy) activates the epidermal growth factor receptor (EGFR), a major effector of the p42/44 mitogen-activated protein kinase (MAPK) pathway. MAPK and its downstream effector, p90 ribosomal S6 kinase (p90RSK), phosphorylate transcription factors involved in cell proliferation. To establish the role of the EGFR/MAPK pathway in radiation-induced transcription factor activation, MDA-MB-231 human breast carcinoma cells were examined using specific inhibitors of signaling pathways. Gel-shift analysis revealed three different profile groups: 1) transcription factors that responded to both radiation (2 Gy) and epidermal growth factor (EGF) (CREB, Egr, Ets, and Stat3); 2) factors that responded to radiation, but not EGF (C/EBP and Stat1); and 3) those that did not respond significantly to either radiation or EGF (AP-1 and Myc). Within groups 1 and 2, a two- to fivefold maximum stimulation of binding activity was observed at 30–60 min after irradiation. Interestingly, only transcription factors that responded to EGF had radiation responses significantly inhibited by the EGFR tyrosine kinase inhibitor, AG1478; these responses were also abrogated by farnesyltransferase inhibitor (FTI) or PD98059, inhibitors of Ras and MEK1/2, respectively. Moreover, radiation-induced increases in CREB and p90RSK phosphorylation and activation of Stat3 and Egr-1 reporter constructs by radiation were all abolished by AG1478. These data demonstrate a distinct radiation response profile at the transcriptional level that is dependent on enhanced EGFR/Ras/MAPK signaling

Studying the Solar system with the International Pulsar Timing Array

Pulsar-timing analyses are sensitive to errors in the Solar-system ephemerides (SSEs) that timing models utilize to estimate the location of the Solar-system barycentre, the quasi-inertial reference frame to which all recorded pulse times-of-arrival are referred. Any error in the SSE will affect all pulsars, therefore pulsar timing arrays (PTAs) are a suitable tool to search for such errors and impose independent constraints on relevant physical parameters. We employ the first data release of the International Pulsar Timing Array to constrain the masses of the planet-moons systems and to search for possible unmodelled objects (UMOs) in the Solar system. We employ 10 SSEs from two independent research groups, derive and compare mass constraints of planetary systems, and derive the first PTA mass constraints on asteroidbelt objects. Constraints on planetary-system masses have been improved by factors of up to 20 from the previous relevant study using the same assumptions, with the mass of the Jovian system measured at 9.5479189(3) × 10 -4 M ⊙ . The mass of the dwarf planet Ceres is measured at 4.7(4) × 10 -10 M ⊙ . We also present the first sensitivity curves using real data that place generic limits on the masses of UMOs, which can also be used as upper limits on the mass of putative exotic objects. For example, upper limits on dark-matter clumps are comparable to published limits using independent methods.While the constraints on planetary masses derived with all employed SSEs are consistent, we note and discuss differences in the associated timing residuals and UMO sensitivity curves

Clinical and Molecular Phenotype of Aicardi-Goutières Syndrome

Aicardi-Goutières syndrome (AGS) is a genetic encephalopathy whose clinical features mimic those of acquired in utero viral infection. AGS exhibits locus heterogeneity, with mutations identified in genes encoding the 3′→5′ exonuclease TREX1 and the three subunits of the RNASEH2 endonuclease complex. To define the molecular spectrum of AGS, we performed mutation screening in patients, from 127 pedigrees, with a clinical diagnosis of the disease. Biallelic mutations in TREX1, RNASEH2A, RNASEH2B, and RNASEH2C were observed in 31, 3, 47, and 18 families, respectively. In five families, we identified an RNASEH2A or RNASEH2B mutation on one allele only. In one child, the disease occurred because of a de novo heterozygous TREX1 mutation. In 22 families, no mutations were found. Null mutations were common in TREX1, although a specific missense mutation was observed frequently in patients from northern Europe. Almost all mutations in RNASEH2A, RNASEH2B, and RNASEH2C were missense. We identified an RNASEH2C founder mutation in 13 Pakistani families. We also collected clinical data from 123 mutation-positive patients. Two clinical presentations could be delineated: an early-onset neonatal form, highly reminiscent of congenital infection seen particularly with TREX1 mutations, and a later-onset presentation, sometimes occurring after several months of normal development and occasionally associated with remarkably preserved neurological function, most frequently due to RNASEH2B mutations. Mortality was correlated with genotype; 34.3% of patients with TREX1, RNASEH2A, and RNASEH2C mutations versus 8.0% RNASEH2B mutation–positive patients were known to have died (P=.001). Our analysis defines the phenotypic spectrum of AGS and suggests a coherent mutation-screening strategy in this heterogeneous disorder. Additionally, our data indicate that at least one further AGS-causing gene remains to be identified