Oncogenic Ras upregulates NADPH oxidase 1 gene expression through MEK-ERK-dependent phosphorylation of GATA-6.

Ras oncogene upregulates the expression of nicotinamide adenine dinucleotide phosphate oxidase (Nox) 1 via the Raf/MEK/ERK pathway, leading to the elevated production of reactive oxygen species that is essential for maintenance of Ras-transformation phenotypes. However, the precise transcriptional control mechanism underlying Ras-induced Nox1 expression remains to be elucidated. Here we demonstrated that via the MEK/ERK pathway, Ras signaling enhances the activity of the functional Nox1 promoter (nt -321 to -1) in colon cancer CaCo-2 cells and thereby induces the formation of the specific protein-DNA complexes in the two GATA-binding site-containing regions (nt -161 to -136 and -125 to -100). Supershift assays with GATA antibodies, protein analyses and chromatin immunoprecipitation revealed that GATA-6 is a component of the specific protein-DNA complexes at the Nox1 promoter. GATA-6 was able to trans-activate the Nox1 promoter but not a promoter in which the GATA-binding sites are mutated. Moreover, GATA-6 was phosphorylated at serine residues by MEK-activated ERK, which increased GATA-6 DNA binding, correlating with suppression of the Nox1 promoter activity by an MEK inhibitor PD98059. Finally, the site-directed mutation of the consensus ERK phosphorylation site (PYS(120)P to PYA(120)P) of GATA-6 abolished its trans-activation activity, suppressing of the growth of CaCo-2 cells. On the basis of these results, we propose that oncogenic Ras signaling upregulates the transcription of Nox1 through MEK-ERK-dependent phosphorylation of GATA-6.

XANES spectra of mechanical alloyed Y-Cu system.

We report changes in the structure and the electronic states of YCu2 during mechanical alloying. The x-ray diffraction shows that milling for about 60 h transforms mixture of elemental powders of Y and Cu into an amorphous alloy. We have measured the x-ray absorption near edge structure (XANES) spectra of mechanically alloyed YCu2 at Cu K and Y K edges at various milling time. The pre-edge peak near the Cu K threshold grows remarkably and its central energy decreases with increasing the milling time. The absorption near the Y K threshold, on the other hand, decreases with milling time. We discuss these features of XANES spectra in connection with the phase transformation from the crystalline to amorphous states.

XANES study in the solid state reaction of Zr-Ni system.

We have measured x-ray absorption near edge structure (XANES) spectra of mechanically alloyed ZrNi near Ni and Zr K edges as a function of milling time. The x-ray diffraction shows that the system is transformed into an amorphous phase by milling for about 60 h. With increasing the milling time, a small absorption near the Ni threshold rises and finally forms a peak in the amorphous phase, while an absorption near the Zr threshold shows a gradual decrease. The peak of the Ni K edge reflects 3d character strongly hybridized with 4p states, while the absorption of the Zr K edge comes from states having a 4d orbital character.