The PML tumor suppressor controls key pathways for growth suppression, induction of apoptosis, and cellular senescence. PML loss occurs frequently in human tumors through unknown posttranslational mechanisms. Casein kinase 2 (CK2) is oncogenic and frequently upregulated in human tumors. Here we show that CK2 regulates PML protein levels by promoting its ubiquitin-mediated degradation dependent on direct phosphorylation at Ser517. Consequently, PML mutants that are resistant to CK2 phosphorylation display increased tumor-suppressive functions. In a faithful mouse model of lung cancer, we demonstrate that Pml inactivation leads to increased tumorigenesis. Furthermore, CK2 pharmacological inhibition enhances the PML tumor-suppressive property in vivo. Importantly, we found an inverse correlation between CK2 kinase activity and PML protein levels in human lung cancer-derived cell lines and primary specimens. These data identify a key posttranslational mechanism that controls PML protein levels and provide therapeutic means toward PML restoration through CK2 inhibition.

IntroductionLeft untreated, malignant pleural mesothelioma (MPM) is associated with uniformly poor prognosis. Better survival has been reported with surgery-based multimodality therapy, but to date, no trial has demonstrated survival benefit of surgery over other therapies. We evaluated whether cancer-directed surgery influenced survival independently from other predictors in a large population-based dataset.MethodsThe SEER database was explored from 1973 to 2009 to identify all cases of pathologically-proven MPM. Age, sex, race, year of diagnosis, histology stage, cancer-directed surgery, radiation, and vital status were analyzed. The association between prognostic factors and survival was estimated using Cox regression and propensity matched analysis.ResultsThere were 14,228 patients with pathologic diagnosis of MPM. On multivariable analysis, female gender, younger age, early stage, and treatment with surgery were independent predictors of longer survival. In comparison to no treatment, surgery alone was associated with significant improvement in survival [adjusted hazard ratio (adj HR) 0.64 (0.61–0.67)], but not radiation [adj HR 1.15 (1.08–1.23)]. Surgery and radiation combined had similar survival as surgery alone [adj HR 0.69 (0.64–0.76)]. Results were similar when cases diagnosed between 1973 and 1999 were compared to cases diagnosed between 2000 and 2009.ConclusionsDespite developments in surgical and radiation techniques, the prognosis for MPM patients has not improved over the past 4 decades. Cancer-directed surgery is independently associated with better survival, suggesting that multimodal surgery-based therapy can benefit these patients. Further research in adjuvant treatment is necessary to improve prognosis in this challenging disease.

Chromosomal amplification at 3q is common to multiple human cancers, but has a specific predilection for squamous cell carcinomas (SCC) of mucosal origin. We identified and characterized a novel oncogene, SCC-related oncogene (SCCRO), which is amplified along the 3q26.3 region in human SCC. Amplification and overexpression of SCCRO in these tumors correlate with poor clinical outcome. The importance of SCCRO amplification in malignant transformation is established by the apoptotic response to short hairpin RNA against SCCRO, exclusively in cancer cell lines carrying SCCRO amplification. The oncogenic potential of SCCRO is underscored by its ability to transform fibroblasts (NIH-3T3 cells) in vitro and in vivo. We show that SCCRO regulates Gli1-a key regulator of the hedgehog (HH) pathway. Collectively, these data suggest that SCCRO is a novel component of the HH signaling pathway involved in the malignant transformation of squamous cell lineage.

Covalent modification of cullins by the ubiquitin-like protein NEDD8 (neddylation) regulates protein ubiquitination by promoting the assembly of cullin-RING ligase E3 complexes. Like ubiquitination, neddylation results from an enzymatic cascade involving the sequential activity of a dedicated E1 (APPBP1/ Uba3), E2 (Ubc12), and an ill-defined E3. We show that SCCRO (also known as DCUN1D1) binds to the components of the neddylation pathway (Cullin-ROC1, Ubc12, and CAND1) and augments but is not required for cullin neddylation in reactions using purified recombinant proteins. We also show that SCCRO recruits Ubc12ϳNEDD8 to the CAND1-Cul1-ROC1 complex but that this is not sufficient to dissociate or overcome the inhibitory effects of CAND1 on cullin neddylation in purified protein assays. In contrast to findings in cellular systems where no binding is seen, we show that SCCRO and CAND1 can bind to the neddylated Cul1-ROC1 complex in assays using purified recombinant proteins. Although neddylated (not unneddylated) Cul1-ROC1 is released from CAND1 upon incubation with testis lysate from SCCRO ؉/؉ mice, the addition of recombinant SCCRO is required to achieve the same results in lysate from SCCRO ؊/؊ mice. Combined, these results suggest that SCCRO is an important component of the neddylation E3 complex that functions to recruit charged E2 and is involved in the release of inhibitory effects of CAND1 on cullin-RING ligase E3 complex assembly and activity.Post-translational modification of proteins by ubiquitin (Ub) 4 regulates diverse cellular functions including protein turnover, differentiation, apoptosis, cell cycle, and transcription (1-5). Given its essential role, ubiquitination is a highly regulated process that involves the sequential action of three enzymes termed as E1, E2, and E3. In this enzymatic cascade, E1 initiates the process by forming a high energy thioester bond with Ub in an ATP-coupled reaction. The Ub is then transferred to E2 as a thioester intermediate. Finally, E3s serve as the targeting arm in the ubiquitination process, mediating the transfer of Ub from E2 to the target protein to create an isopeptide bond between the C-terminal glycine in Ub and a lysine residue on the substrate protein. Once attached, the Ub itself can be modified to generate polyubiquitin chains on the target protein (6). The functional effects of ubiquitination are influenced by the chain length and the residue on the Ub to which the chain is attached. Polyubiquitination promotes translocation to the 26 S proteasome for degradation. Other functional effects of mono-and polyubiquitination include protein translocation, interaction, and activation.Although there is only one known E1 (except in plants) and relatively few E2s, E3s exist in multiple forms to allow for specific protein targeting (6). In general, E3s are modular multiprotein complexes that can be divided into two broad categories based on the presence of either a HECT (homologous to E6-AP C terminus) or RING (Really Interesting New Gene)-finger domain-containi...

SCCRO/DCUN1D1/DCN1 (squamous cell carcinoma-related oncogene/defective in cullin neddylation 1 domain containing 1/defective in cullin neddylation) serves as an accessory E3 in neddylation by binding to cullin and Ubc12 to allow efficient transfer of Nedd8. In this work we show that SCCRO has broader, pleiotropic effects that are essential for cullin neddylation in vivo. Reduced primary nuclear localization of Cul1 accompanying decreased neddylation and proliferation in SCCRO ؊/؊ mouse embryonic fibroblasts led us to investigate whether compartmentalization plays a regulatory role. Decreased nuclear localization, neddylation, and defective proliferation in SCCRO ؊/؊ mouse embryonic fibroblasts were rescued by transgenic expression of SCCRO. Expression of reciprocal SCCRO and Cul1-binding mutants confirmed the requirement for SCCRO in nuclear translocation and neddylation of cullins in vivo. Nuclear translocation of Cul1 by tagging with a nuclear localization sequence allowed neddylation independent of SCCRO, but at a lower level. We found that in the nucleus, SCCRO enhances recruitment of Ubc12 to Cul1 to promote neddylation. These findings suggest that SCCRO has an essential role in neddylation in vivo involving nuclear localization of neddylation components and recruitment and proper positioning of Ubc12.The cullin family of proteins anchor Cullin RING finger-type E3 ubiquitination complexes (CRL) 4 that regulate the degradation and activity of proteins involved in a wide range of cellular processes (1, 2). Several reports have shown that the activity of CRL complexes is primarily regulated by neddylation, a process mechanistically analogous to ubiquitination, where the cullin family of proteins are covalently modified by ubiquitin (Ub)-like protein Nedd8 (3-11). All cullin proteins in humans (Cul1, Cul2, Cul3, Cul4a, Cul4b, Cul5, and PARC) are subject to neddylation, with the exception of Cul7 (12, 13). Lethality resulting from knocking out core components in all organisms studied (except budding yeast) emphasizes the indispensable role of neddylation in normal cellular function (14 -20).Like ubiquitination, neddylation involves a sequential, tripartite enzymatic cascade (21, 22). The vertebrate enzymes for neddylation are the heterodimeric complex APP-BP1/Uba3 (E1) and Ubc12 or Ube2f (E2) (21,(23)(24)(25). The presence of enzymatic activity in the RING domain of Roc1 combined with its requirement and sufficiency to promote Nedd8 conjugation in vitro supports a role for Roc1 as the E3 for neddylation (26). Recent work identified a novel protein SCCRO/DCUN1D1/ DCN1 (squamous cell carcinoma-related oncogene/defective in cullin neddylation 1, domain containing 1/defective in cullin neddylation) that binds to components of the E3 complex for neddylation (Ubc12 and Cullin-Roc1) and increases neddylation efficiency in vitro (27)(28)(29)(30). Biochemical studies and structural modeling suggest that DCN1 functions as an E3 promoting neddylation by reducing nonspecific Rub1 (Nedd8 in mammals) discharge and directing...

Figure 1: Pipeline: The user specifies the model and its corresponding animation. Our system computes the required activations, and simulates the muscles, tendons, and bones. The skin is then attached to the skeleton, and the subcutaneous deformation from tendon motion is added as a post-process. AbstractWe describe an automatic technique for generating the motion of tendons and muscles under the skin of a traditionally animated character. This is achieved by integrating the traditional animation pipeline with a novel biomechanical simulator capable of dynamic simulation with complex routing constraints on muscles and tendons. We also describe an algorithm for computing the activation levels of muscles required to track the input animation. We demonstrate the results with several animations of the human hand.

Introduction: The TNM classification for lung cancer, originally designed for NSCLC, is applied to staging of bronchopulmonary carcinoid tumors. The validity of the eighth edition of the staging system for carcinoid tumors has not been assessed. In this study, we evaluated its prognostic accuracy by using data from a large national population-based cancer registry.Methods: Patients with typical and atypical bronchopulmonary carcinoids diagnosed between 2000 and 2013 were identified from the National Cancer Institute's Surveillance, Epidemiology and End Results registry. We used competing risks analysis to compare 10-year diseasespecific survival (DSS) across stages.Results: Overall, 4645 patients with bronchopulmonary carcinoid tumors were identified. Worsening DSS with increasing TNM status and stage was demonstrated across both typical and atypical carcinoids, with overlaps between adjacent subcategories. The combined stages (I versus II, II versus III, and III versus IV) showed greater separation in DSS despite persistent overlaps between groups. For typical carcinoids, we found decreased DSS for stages II, III, and IV, with hazard ratios of 3.8 (95% confidence interval [CI]: 2.6-5.6), 4.3 (95% CI: 3.0-6.1), and 9.0 (95% CI: 6.1-13.1), respectively, compared with stage I. Conclusion:The combined stage categories of the eighth edition of the TNM staging system provide useful information on outcomes for typical and atypical carcinoids. However, persistent overlaps in combined stage and subcategories of the staging system limit the usefulness of the TNM staging system, particularly in intermediate stages. These limitations suggest the need for future further study and refinement.

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