S-Nitrosylation at Cysteine 498 of c-Src Tyrosine Kinase Regulates Nitric Oxide-mediated Cell Invasion

• Published in: The Journal of biological chemistry 2010-02, Vol.285 (6), p.3806-3814
• Main Authors: Rahman, Mohammad Aminur, Senga, Takeshi, Ito, Satoko, Hyodo, Toshinori, Hasegawa, Hitoki, Hamaguchi, Michinari
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Blotting, Western; Breast Cancer; Cadherins - metabolism; Cell Adhesion - drug effects; Cell Line, Tumor; Cell Movement; Cell/Migration; Cells, Cultured; Cysteine - metabolism; Estradiol - pharmacology; Humans; Mechanisms of Signal Transduction; Mice; Mice, Knockout; Microscopy, Fluorescence; Molecular Sequence Data; Mutation; Nitric Oxide; Nitric Oxide - metabolism; Nitric Oxide Donors - pharmacology; Nitrosation - drug effects; Proto-Oncogene Proteins c-fyn - genetics; Proto-Oncogene Proteins c-fyn - metabolism; Proto-Oncogene Proteins c-yes - genetics; Proto-Oncogene Proteins c-yes - metabolism; RNA Interference; S-Nitroso-N-Acetylpenicillamine - pharmacology; S-nitrosylation; Sequence Homology, Amino Acid; Signal Transduction; Signal Transduction/Phosphotyrosine; Signal Transduction/Protein Kinases/Tyrosine; Src; src-Family Kinases - genetics; src-Family Kinases - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M109.059782

Nitric oxide (NO) plays a pivotal role in tumorigenesis, particularly with relation to cancer cell invasion and metastasis. NO can reversibly couple to cysteine thiols to form an S-nitrosothiol, which regulates the enzymatic activities of target proteins. c-Src is a tyrosine kinase that promotes cancer cell invasion and metastasis. Interestingly, c-Src can be activated by NO stimulation. However, mechanisms by which NO stimulates Src kinase activity have not been elucidated. We report here that NO causes S-nitrosylation of c-Src at cysteine 498 (Cys498) to stimulate its kinase activity. Cys498 is conserved among Src family kinases, and Cys506 of c-Yes, which corresponds to Cys498 of c-Src, was also important for the NO-mediated activation of c-Yes. Estrogens may work synergistically with NO to induce the proliferation and migration of many kinds of breast cancer cells. For example, β-estradiol induces the expression of endothelial nitric synthase and production of NO in MCF7 cells. We found that activation of c-Src in MCF7 cells by β-estradiol stimulation was mediated by the S-nitrosylation of Cys498. In addition, we report that disruption of E-cadherin junctions and enhancement of cell invasion by β-estradiol stimulation was mediated by NO-dependent activation of c-Src. These results identify a novel signaling pathway that links NO and Src family kinases to cancer cell invasion and metastasis.