ANIA:ANnotation and Integrated Analysis of the 14-3-3 interactome

The dimeric 14-3-3 proteins dock onto pairs of phosphorylated Ser and Thr residues on hundreds of proteins, and thereby regulate many events in mammalian cells. To facilitate global analyses of these interactions, we developed a web resource named ANIA: ANnotation and Integrated Analysis of the 14-3-3 interactome, which integrates multiple data sets on 14-3-3-binding phosphoproteins. ANIA also pinpoints candidate 14-3-3-binding phosphosites using predictor algorithms, assisted by our recent discovery that the human 14-3-3-interactome is highly enriched in 2R-ohnologues. 2R-ohnologues are proteins in families of two to four, generated by two rounds of whole genome duplication at the origin of the vertebrate animals. ANIA identifies candidate ‘lynchpins’, which are 14-3-3-binding phosphosites that are conserved across members of a given 2R-ohnologue protein family. Other features of ANIA include a link to the catalogue of somatic mutations in cancer database to find cancer polymorphisms that map to 14-3-3-binding phosphosites, which would be expected to interfere with 14-3-3 interactions. We used ANIA to map known and candidate 14-3-3-binding enzymes within the 2R-ohnologue complement of the human kinome. Our projections indicate that 14-3-3s dock onto many more human kinases than has been realized. Guided by ANIA, PAK4, 6 and 7 (p21-activated kinases 4, 6 and 7) were experimentally validated as a 2R-ohnologue family of 14-3-3-binding phosphoproteins. PAK4 binding to 14-3-3 is stimulated by phorbol ester, and involves the ‘lynchpin’ site phosphoSer99 and a major contribution from Ser181. In contrast, PAK6 and PAK7 display strong phorbol ester-independent binding to 14-3-3, with Ser113 critical for the interaction with PAK6. These data point to differential 14-3-3 regulation of PAKs in control of cell morphology. Database URL: https://ania-1433.lifesci.dundee.ac.uk/prediction/webserver/index.p

Corneal Distortion Eliminated by Refitting with Gas Permeable Lenses

Corneal Distortion Eliminated by Refitting with Gas Permeable Lense

NA

http://www.archive.org/details/somecreepfractur00molzU.S. Navy (U.S.N.) author

Synergistic binding of the phosphorylated S233- and S259-binding sites of C-RAF to one 14-3-3ζ dimer

C-RAF kinase is a central component of the Ras-RAF-MEK (mitogen-activated protein kinase/extracellular signal-regulated kinase)-ERK (extracellular signal-regulated kinase) pathway, which has been shown to be activated in 30% of human tumors. 14-3-3 proteins inactivate C-RAF by binding to the two N-terminal phosphorylation-dependent binding sites surrounding S233 and S259. 14-3-3 proteins can bind two target sequences located on one polypeptide chain simultaneously, thereby increasing binding affinity compared to single-site binding and possibly allowing regulated 14-3-3 binding through gatekeeper phosphorylation. To date, it was unclear whether 14-3-3 proteins can bind the two N-terminal phosphorylation-dependent binding sites of C-RAF simultaneously. Fluorescence polarization using phosphorylated peptides demonstrated that S233 is the low-affinity and S259 is the high-affinity binding site, while simultaneous engagement of both sites by 14-3-3¿ enhances affinity compared to single-site binding. Determination of a 1:1 stoichiometry for the di-phosphorylated peptide binding to one 14-3-3¿ dimer with isothermal titration calorimetry was supported by the crystal structure of the 14-3-3¿/C-RAFpS233,pS259 complex. Cellular localization studies validate the significance of these sites for cytoplasmic retention of C-RAF, suggesting an extended mechanism of RAF regulation by 14-3-3 proteins

Education for expectant fathers in workplaces in Turkey

Worldwide, there is increasing recognition that if family and reproductive health programmes are to be successful, the involvement of men is essential. As part of the problem, men also have to be seen as part of the solution. The reality is that in many countries, including Turkey, men generally do not accompany their partners to health facilities for family planning, antenatal and postnatal services and are not expected to attend the labour or birth of their child. Workplace programmes are a potential strategy for meeting the reproductive health education needs of men in industrial cities such as Istanbul. This intervention study was developed to test the feasibility and effects of expanding a special programme for expectant fathers to large workplaces in Istanbul, with the aim of improving the health of Turkish families during the pregnancy, birth and newborn periods. The findings indicate that it is possible to train workplace physicians in Istanbul to conduct regular educational programmes for expectant fathers on reproductive health, and that such programmes may have beneficial effects, especially in the areas of pregnancy nutrition, exclusive breast-feeding, and support behaviours. Considering the difficulty of getting men to attend hospital or clinic-based educational programmes in large urban areas, bringing such training programmes to men at their places of work has the potential to be an important strategy. Given that large workplaces in Turkey already have full-time physicians charged with the duty of health education for employees, this is also a feasible strategy