Molecular genetic structure-function analysis of translation initiation factor eIF5B.

摘要: Recently, significant progress has been made in obtaining three‐dimensional (3‐D) structures of the factors that promote translation initiation, elongation, and termination. These structures, when interpreted light previous biochemical characterizations factors, provide insight into function molecular mechanism specific steps process. In addition, genetic analyses yeast have helped elucidate vivo roles various pathway. We combined these two approaches use studies to define structure–function properties initiation Saccharomyces cerevisiae. this chapter, we describe our multistep approach which first characterize a site‐directed mutant factor interest using vitro assays protein synthesis. Next, subject gene random mutagenesis screen for second‐site mutations restore factor's vivo. Following characterization suppressor mutant, interpret results 3‐D structure new insights translation.