Recombinant adenoviral expression of dominant-negative ras N17 blocking radiation-induced activation of mitogen-activated protein kinase pathway

Publisher Summary This chapter focuses on the recombinant adenoviral expression of dominant-negative Ras N 17 blocking radiation-induced activation of mitogen-activated protein kinase pathway. Generation of recombinant adenoviruses to express a variety of proteins has become a method of choice for many investigators in the field of signal transduction. Using this technique, near 100% infection efficiencies can be obtained. A novel cellular target for ionizing radiation is shown to be the epidermal growth factor receptor (EGFR, also called ErbB 1), which is activated in response to irradiation of several carcinoma cell types. Radiation exposure, via the EGFR, can activate the extracellular signal-regulated kinase-mitogen-activated protein kinase (ERK/MAPK) pathway to a level similar to that observed by physiologic EGF concentrations. The data presented in this chapter describe the generation of a recombinant adenovirus to express dominant-negative Ras N 17 and the effect this gene product has on radiation-induced ERK-MAPK activity.