We present extensive optical (U BV RI), near-infrared (JK) light curves and optical spectroscopy of the Type Ia supernova (SN) 2006X in the nearby galaxy NGC 4321 (M100). Our observations suggest that either SN 2006X has an intrinsically peculiar color evolution, or it is highly reddened [E(B − V ) host = 1.42 ± 0.04 mag] with R V = 1.48 ± 0.06, much lower than the canonical value of 3.1 for the average Galactic dust. SN 2006X also has one of the highest expansion velocities ever published for a SN Ia. Compared with the other SNe Ia we analyzed, SN 2006X has a broader light curve in the U band, a more prominent bump/shoulder feature in the V and R bands, a more pronounced secondary maximum in the I and near-infrared bands, and a remarkably smaller late-time decline rate in the B band. The B − V color evolution shows an obvious deviation from the Lira-Phillips relation at 1 to 3 months after maximum brightness. At early times, optical spectra of SN 2006X displayed strong, high-velocity features of both intermediate-mass elements (Si, Ca, and S) and iron-peak elements, while at late times they showed a relatively blue continuum, consistent with the blue U − B and B − V colors at similar epochs. A light echo and/or the interaction of the SN ejecta and its circumstellar material may provide a plausible explanation for its late-time photometric and spectroscopic behavior. Using the Cepheid distance of M100, we derive a Hubble constant of 72.8 ± 8.2 km s −1 Mpc −1 (statistical) from the normalized dereddened luminosity of SN 2006X. We briefly discuss whether abnormal dust is a universal signature for all SNe Ia, and whether the most rapidly expanding objects form a subclass with distinct photometric and spectroscopic properties.

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Toll-like receptor 4 (TLR4) plays a pivotal role in innate immune responses, and the transcription factor CCAAT/enhancer binding protein delta (C/EBPδ, Cebpd) is a TLR4-induced gene. Here, we identify a positive feedback loop in which C/EBPδ activates Tlr4 gene expression in macrophages and tumour cells. In addition, we discovered a negative feedback loop whereby the tumour suppressor FBXW7α (FBW7, Cdc4), whose gene expression is inhibited by C/EBPδ, targets C/EBPδ for degradation when C/EBPδ is phosphorylated by GSK-3β. Consequently, FBXW7α suppresses Tlr4 expression and responses to the ligand lipopolysaccharide (LPS). FBXW7α depletion alone is sufficient to augment pro-inflammatory signalling in vivo. Moreover, as inflammatory pathways are known to modulate tumour biology, Cebpd null mammary tumours, which have reduced metastatic potential, show altered expression of inflammation-associated genes. Together, these findings reveal a role for C/EBPδ upstream of TLR4 signalling and uncover a function for FBXW7α as an attenuator of inflammatory signalling.

AML1 (RUNX1) regulates hematopoiesis, angiogenesis, muscle function, and neurogenesis. Previous studies have shown that phosphorylation of AML1, particularly at serines 276 and 303, affects its transcriptional activation. Here, we report that phosphorylation of AML1 serines 276 and 303 can be blocked in vivo by inhibitors of the cyclin-dependent kinases (CDKs) Cdk1 and Cdk2. Furthermore, these residues can be phosphorylated in vitro by purified Cdk1/cyclin B and Cdk2/cyclin A. Mutant AML1 protein which cannot be phosphorylated at these sites (AML1-4A) is more stable than wild-type AML1. AML-4A is resistant to degradation mediated by Cdc20, one of the substrate-targeting subunits of the anaphase-promoting complex (APC). However, Cdh1, another targeting subunit used by the APC, can mediate the degradation of AML1-4A. A phospho-mimic protein, AML1-4D, can be targeted by Cdc20 or Cdh1. These observations suggest that both Cdc20 and Cdh1 can target AML1 for degradation by the APC but that AML1 phosphorylation may affect degradation mediated by Cdc20-APC to a greater degree.The AML1 proteins, including AML1a, AML1b, and AML1c (AML1c is also known as AML1B), are generated from one gene by alternative splicing (22). This gene has been given the names RUNX1, AML1, CBFA2, and PEBP2␣B (37). The AML1 protein is composed of a DNA binding runt homology domain located in the amino terminus followed by a transcriptional activation domain and a negative regulatory C-terminal domain (15,20,29). AML1 was initially identified during the study of breakpoint t(8;21), which is a common chromosomal translocation in acute myeloid leukemia (23). The association of AML1 with blood cell development is shown by the disruption of the AML1 gene through multiple chromosomal translocations, deletions, point mutations, or amplification in approximately 30% of human myeloid leukemias and myelodysplastic syndrome patients and a significant number of lymphoid leukemia patients (21,24,27,32). Furthermore, no detectable definitive hematopoiesis is observed in Aml1 knockout mice (26,40). The importance of AML1 in nonhematopoietic cells has also been recognized in angiogenesis, muscle function, and neurogenesis (6,12,14,35,41).AML1 is detected as a serine and threonine phosphorylated protein (9). Previous work has suggested that AML1 activity may be regulated by phosphorylation (34,45). Phosphorylation at specific serine-proline or threonine-proline sites in AML1 appears to be necessary for normal activity (45). It has also been suggested that phosphorylation releases AML1 from an association with the nuclear matrix mediated by sin3A, in turn leading to both increased activity and an increased rate of degradation (11).We have now shown that AML1 phosphorylation by cyclindependent kinases (CDKs) affects the overall stability of AML1 as well as the ability of certain ubiquitin ligase complexes, such as Cdc20-anaphase-promoting complex (APC), to target AML1 for degradation. MATERIALS AND METHODSCell culture and treatment. 293T and NIH 3T3 cells were grown...

The transcription factor CCAAT/enhancer binding protein δ (C/EBPδ, CEBPD, NFIL-6β) has tumor suppressor function; however, the molecular mechanism(s) by which C/EBPδ exerts its effect are largely unknown. Here, we report that C/EBPδ induces expression of the Cdc27 (APC3) subunit of the anaphase promoting complex/cyclosome (APC/C), which results in the polyubiquitination and degradation of the prooncogenic cell cycle regulator cyclin D1, and also down-regulates cyclin B1, Skp2, and Plk-1. In C/EBPδ knockout mouse embryo fibroblasts (MEF) Cdc27 levels were reduced, whereas cyclin D1 levels were increased even in the presence of activated GSK-3β. Silencing of C/EBPδ, Cdc27, or the APC/C coactivator Cdh1 (FZR1) in MCF-10A breast epithelial cells increased cyclin D1 protein expression. Like C/EBPδ, and in contrast to cyclin D1, Cdc27 was down-regulated in several breast cancer cell lines, suggesting that Cdc27 itself may be a tumor suppressor. Cyclin D1 is a known substrate of polyubiquitination complex SKP1/CUL1/F-box (SCF), and our studies show that Cdc27 directs cyclin D1 to alternative degradation by APC/C. These findings shed light on the role and regulation of APC/C, which is critical for most cellular processes.

We find that phorbol ester (PE) treatment of K562 cells greatly stimulates promoters (T cell receptor ␤, myeloperoxidase, macrophage colony-stimulating factor receptor, and granulocyte macrophage colony-stimulating factor receptor) containing AML1 transcription factor binding sites. This stimulation of AML1c transcriptional activity is mediated by direct phosphorylation of the AML1c molecule on multiple phosphorylation sites. Eleven AML1c (S/T)P sites in the transcriptional activating domain are phosphorylated at a basal level in untreated K562 cells; treatment of the K562 cells with PE results in increased phosphorylation at five of these sites (serines 276, 293, 303, 462, and threonine 300). Mutation of these five sites to alanine inhibits PE-induced transcriptional activity; mutation of the sites to an acidic amino acid, aspartic acid, stimulates constitutive activity. Single mutations in four amino acids or double mutations (serines 276 and 293 or threonine 300 and serine 303) have little effect on AML1c transcriptional activity. Inhibitor assays suggest that the ERK family of protein kinases is activated by PEs to phosphorylate the (S/T)P sites within the AML1c molecule and markedly enhance the transcriptional activity of AML1c.

Notch signaling represents a key mechanism mediating cancer metastasis and stemness. To understand how Notch signaling is overactivated to couple tumor metastasis and self-renewal in NSCLC cells, we performed the current study and showed that RFC4, a DNA replication factor amplified in more than 40% of NSCLC tissues, directly binds to the Notch1 intracellular domain (NICD1) to competitively abrogate CDK8/FBXW7-mediated degradation of NICD1. Moreover, RFC4 is a functional transcriptional target gene of Notch1 signaling, forming a positive feedback loop between high RFC4 and NICD1 levels and sustained overactivation of Notch signaling, which not only leads to NSCLC tumorigenicity and metastasis but also confers NSCLC cell resistance to treatment with the clinically tested drug DAPT against NICD1 synthesis. Furthermore, together with our study, analysis of two public datasets involving more than 1500 NSCLC patients showed that RFC4 gene amplification, and high RFC4 and NICD1 levels were tightly correlated with NSCLC metastasis, progression and poor patient prognosis. Therefore, our study characterizes the pivotal roles of the positive feedback loop between RFC4 and NICD1 in coupling NSCLC metastasis and stemness properties and suggests its therapeutic and diagnostic/prognostic potential for NSCLC therapy.

In this study, a loop-mediated isothermal amplification (LAMP) method to rapidly detect Staphylococcus aureus strains was developed and evaluated by extensively applying a large number of S. aureus isolates from clinical and food samples. Six primers were specially designed for recognizing eight distinct sequences on the species-specific femA gene of S. aureus. The detection limits were 100 fg DNA/tube and 10(4) CFU/ml. The LAMP assay was applied to 432 S. aureus strains isolated from 118 clinical and 314 food samples. Total detection rates for the LAMP and polymerase chain reaction assays were 98.4% (306/311) and 89.4% (278/311), respectively.