Tumorigenesis results from dysregulation of oncogenes and tumor suppressors that influence cellular proliferation, differentiation, apoptosis, and/or senescence. Many gene products involved in these ...processes are substrates of the E3 ubiquitin ligase Mule/Huwe1/Arf-BP1 (Mule), but whether Mule acts as an oncogene or tumor suppressor in vivo remains controversial. We generated K14Cre;Mule(flox/flox(y)) (Mule kKO) mice and subjected them to DMBA/PMA-induced skin carcinogenesis, which depends on oncogenic Ras signaling. Mule deficiency resulted in increased penetrance, number, and severity of skin tumors, which could be reversed by concomitant genetic knockout of c-Myc but not by knockout of p53 or p19Arf. Notably, in the absence of Mule, c-Myc/Miz1 transcriptional complexes accumulated, and levels of p21CDKN1A (p21) and p15INK4B (p15) were down-regulated. In vitro, Mule-deficient primary keratinocytes exhibited increased proliferation that could be reversed by Miz1 knockdown. Transfer of Mule-deficient transformed cells to nude mice resulted in enhanced tumor growth that again could be abrogated by Miz1 knockdown. Our data demonstrate in vivo that Mule suppresses Ras-mediated tumorigenesis by preventing an accumulation of c-Myc/Miz1 complexes that mediates p21 and p15 down-regulation.
Spatiotemporal control of gene expression is central to animal development. Core promoters represent a previously unanticipated regulatory level by interacting with cis-regulatory elements and ...transcription initiation in different physiological and developmental contexts. Here, we provide a first and comprehensive description of the core promoter repertoire and its dynamic use during the development of a vertebrate embryo. By using cap analysis of gene expression (CAGE), we mapped transcription initiation events at single nucleotide resolution across 12 stages of zebrafish development. These CAGE-based transcriptome maps reveal genome-wide rules of core promoter usage, structure, and dynamics, key to understanding the control of gene regulation during vertebrate ontogeny. They revealed the existence of multiple classes of pervasive intra- and intergenic post-transcriptionally processed RNA products and their developmental dynamics. Among these RNAs, we report splice donor site-associated intronic RNA (sRNA) to be specific to genes of the splicing machinery. For the identification of conserved features, we compared the zebrafish data sets to the first CAGE promoter map of Tetraodon and the existing human CAGE data. We show that a number of features, such as promoter type, newly discovered promoter properties such as a specialized purine-rich initiator motif, as well as sRNAs and the genes in which they are detected, are conserved in mammalian and Tetraodon CAGE-defined promoter maps. The zebrafish developmental promoterome represents a powerful resource for studying developmental gene regulation and revealing promoter features shared across vertebrates.
Diverse functions have been reported for lipocalin 2. To investigate these functions in vivo, we generated gene-targeted lipocalin 2-deficient mice ($Lcn2^{-/-}$mice). In vitro studies have suggested ...that lipocalin 2 is important for cellular apoptosis induced by IL-3 withdrawal, and for the induction of kidney differentiation during embryogenesis. Analysis of$Lcn2^{-/-}$mice showed normal cell death upon IL-3 withdrawal and normal kidney development. However, we found that$Lcn2^{-/-}$mice exhibited an increased susceptibility to bacterial infections, in keeping with the proposed function of lipocalin 2 in iron sequestration. Neutrophils isolated from$Lcn2^{-/-}$mice showed significantly less bacteriostatic activity compared with WT controls. The bacteriostatic property of the WT neutrophils was abolished by the addition of exogenous iron, indicating that the main function of lipocalin 2 in the antibacterial innate immune response is to limit this essential element. Another important function ascribed to lipocalin 2 has been its protective role against kidney ischemia-reperfusion injury. We analyzed$Lcn2^{-/-}$mice using a mouse model for severe renal failure and could not detect any significant differences compared with their WT littermates.
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BFBNIB, NMLJ, NUK, PNG, SAZU, UL, UM, UPUK
Phosphorylation of IκB, an inhibitor of
NF-κB
, is an important step in the activation of the transcription factor
NF-κB
. Phosphorylation is mediated by the IκB kinase (IKK) complex, known to ...contain two catalytic subunits:
IKKα
and
IKKβ
. A novel, noncatalytic component of this kinase complex called
NEMO
(
N
F-κB
e
ssential
mo
dulator)/
IKKγ
was identified recently. We have generated
NEMO/IKKγ
-deficient mice by gene targeting. Mutant embryos die at E12.5–E13.0 from severe liver damage due to apoptosis.
NEMO/IKKγ
-deficient primary murine embryonic fibroblasts (MEFs) lack detectable
NF-κB
DNA-binding activity in response to TNFα, IL-1, LPS, and Poly(IC) and do not show stimulus-dependent IκB kinase activity, which correlates with a lack of phosphorylation and degradation of IκBα. Consistent with these data, mutant MEFs show increased sensitivity to TNFα-induced apoptosis. Our data provide in vivo evidence that
NEMO/IKKγ
is the first essential, noncatalytic component of the IKK complex.
Abstract
We have compared the star-formation properties of the W49A and W51 regions by using far-infrared data from the Herschel infrared Galactic Plane Survey (Hi-GAL) and 850-$\mu$m observations ...from the James Clerk Maxwell Telescope (JCMT) to obtain luminosities and masses, respectively, of associated compact sources. The former are infrared luminosities from the catalogue of Elia et al., while the latter are from the JCMT Plane survey source catalogue as well as measurements from new data. The clump-mass distributions of the two regions are found to be consistent with each other, as are the clump-formation efficiency and star-formation efficiency analogues. However, the frequency distributions of the luminosities of the young stellar objects are significantly different. While the luminosity distribution in W51 is consistent with Galaxy-wide samples, that of W49A is top heavy. The differences are not dramatic and are concentrated in the central regions of W49A. However, they suggest that physical conditions there, which are comparable in part to those in extragalactic starbursts, are significantly affecting the star-formation properties or evolution of the dense clumps in the region.
Recurrent 15q13.3 microdeletions were recently identified with identical proximal (BP4) and distal (BP5) breakpoints and associated with mild to moderate mental retardation and epilepsy.
To assess ...further the clinical implications of this novel 15q13.3 microdeletion syndrome, 18 new probands with a deletion were molecularly and clinically characterised. In addition, we evaluated the characteristics of a family with a more proximal deletion between BP3 and BP4. Finally, four patients with a duplication in the BP3-BP4-BP5 region were included in this study to ascertain the clinical significance of duplications in this region.
The 15q13.3 microdeletion in our series was associated with a highly variable intra- and inter-familial phenotype. At least 11 of the 18 deletions identified were inherited. Moreover, 7 of 10 siblings from four different families also had this deletion: one had a mild developmental delay, four had only learning problems during childhood, but functioned well in daily life as adults, whereas the other two had no learning problems at all. In contrast to previous findings, seizures were not a common feature in our series (only 2 of 17 living probands). Three patients with deletions had cardiac defects and deletion of the KLF13 gene, located in the critical region, may contribute to these abnormalities. The limited data from the single family with the more proximal BP3-BP4 deletion suggest this deletion may have little clinical significance. Patients with duplications of the BP3-BP4-BP5 region did not share a recognisable phenotype, but psychiatric disease was noted in 2 of 4 patients.
Overall, our findings broaden the phenotypic spectrum associated with 15q13.3 deletions and suggest that, in some individuals, deletion of 15q13.3 is not sufficient to cause disease. The existence of microdeletion syndromes, associated with an unpredictable and variable phenotypic outcome, will pose the clinician with diagnostic difficulties and challenge the commonly used paradigm in the diagnostic setting that aberrations inherited from a phenotypically normal parent are usually without clinical consequences.
It is well known that protein tyrosine phosphatases (PTPs) that become oxidized due to exposure to reactive oxygen species (ROS) undergo a conformational change and are inactivated. However, whether ...PTPs can actively regulate ROS levels in order to prevent PTP inhibition has yet to be investigated. Here, we demonstrate that PTP non-receptor type 12 (PTPN12) protects cells against aberrant ROS accumulation and death induced by oxidative stress. Murine embryonic fibroblasts (MEFs) deficient in PTPN12 underwent increased ROS-induced apoptosis under conditions of antioxidant depletion. Cells lacking PTPN12 also showed defective activation of FOXO1/3a, transcription factors required for the upregulation of several antioxidant genes. PTPN12-mediated regulation of ROS appeared to be mediated by phosphoinositide-dependent kinase-1 (PDK1), which was hyperstimulated in the absence of PTPN12. As tight regulation of ROS to sustain survival is a key feature of cancer cells, we examined PTPN12 levels in tumors from a cohort of breast cancer patients. Patients whose tumors showed high levels of PTPN12 transcripts had a significantly poorer prognosis. Analysis of tissues from patients with various breast cancer subtypes revealed that more triple-negative breast cancers, the most aggressive breast cancer subtype, showed high PTPN12 expression than any other subtype. Furthermore, both human breast cancer cells and mouse mammary epithelial tumor cells engineered to lack PTPN12 exhibited reduced tumorigenic and metastatic potential in vivo that correlated with their elevated ROS levels. The involvement of PTPN12 in the antioxidant response of breast cancer cells suggests that PTPN12 may represent a novel therapeutic target for this disease.
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DOBA, EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Innovations are being developed with different additives in concrete mixtures to enhance their performance under certain conditions. This work models the split tensile strength through the electrical ...resistivity using a machine learning (ML) algorithm in different concrete mixtures. One of the most frequent nondestructive tests used on concrete is electrical resistivity (Er) for the simplicity of taking measurements on concrete elements. ML involves methods that provide solutions to advanced problems employing computational algorithms. This research employs a support vector regression algorithm, which can predict the split tensile strength value using a nondestructive test. The outcomes of this research exhibit a high correlation between electrical concrete resistivity and split tensile strength over 90%. Support vector regression algorithm achieves accuracy over 93% in the forecasting task. Of note is that the modified concrete contains starch as an addition to prove its performance compared to conventional concrete. Keywords: electrical resistivity; machine learning (ML); support vector regression; tensile strength.
The Neil Gehrels Swift Observatory carried out prompt searches for gravitational-wave (GW) events detected by the LIGO/Virgo Collaboration (LVC) during the second observing run ("O2"). Swift ...performed extensive tiling of eight LVC triggers, two of which had very low false-alarm rates (GW170814 and the epochal GW170817), indicating a high confidence of being astrophysical in origin; the latter was the first GW event to have an electromagnetic counterpart detected. In this paper we describe the follow-up performed during O2 and the results of our searches. No GW electromagnetic counterparts were detected; this result is expected, as GW170817 remained the only astrophysical event containing at least one neutron star after LVC's later retraction of some events. A number of X-ray sources were detected, with the majority of identified sources being active galactic nuclei. We discuss the detection rate of transient X-ray sources and their implications in the O2 tiling searches. Finally, we describe the lessons learned during O2 and how these are being used to improve the Swift follow-up of GW events. In particular, we simulate a population of gamma-ray burst afterglows to evaluate our source ranking system's ability to differentiate them from unrelated and uncataloged X-ray sources. We find that 60%-70% of afterglows whose jets are oriented toward Earth will be given high rank (i.e., "interesting" designation) by the completion of our second follow-up phase (assuming that their location in the sky was observed), but that this fraction can be increased to nearly 100% by performing a third follow-up observation of sources exhibiting fading behavior.
Inhibitor of apoptosis protein (IAP)-binding proteins such as Grim, Reaper and HID have been shown to exert a critical role in regulating caspase activity in species such as D. Melanogaster. However, ...a comparable role for the mammalian homologue of second mitochondrial-derived activator of caspase/direct IAP-binding protein with low pI (Smac/DIABLO) has yet to be clearly established in vivo. Despite tremendous interest in recent years in the use of so-called Smac mimetics to enhance chemotherapeutic potency, our understanding of the true physiologic nature of Smac/DIABLO in regulating programmed cell death (PCD) remains elusive. In order to critically evaluate the role of Smac/DIABLO in regulating mammalian PCD, deficiency of caspase-3 was used as a sensitizing mutation in order to reduce aggregate levels of executioner caspase activity. We observe that combinatorial deletion of Diablo and Casp3, but neither alone, results in perinatal lethality in mice. Consistent with this, examination of both intrinsic and extrinsic forms of PCD in lines of murine embryonic fibroblasts demonstrate that loss of Smac/DIABLO alters both caspase-dependent and caspase-independent intrinsic PCD. Comparative small interfering RNA inhibition studies of X-linked inhibitor of apoptosis, cellular inhibitor of apoptosis (cIAP)-1, cIAP-2, caspase-6 and -7 in both wild-type and Casp3/Diablo DKO mouse embryonic fibroblast lineages, supports a model in which Smac/DIABLO acts to enhance the early phase executioner caspase activity through the modulation of inhibitory interactions between specific IAP family members and executioner caspases-3 and -7.
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DOBA, EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
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