Translation initiation is a complex, multi-step process of fundamental importance in all kingdoms of life, during which the start site of the genetic message transmitted in the form of an RNA molecule (mRNA) is selected, and the level of translation determined. Being the slowest step of protein synthesis, initiation is the phase most often subject to regulation. Here we review, in a historical perspective and focusing mainly on results from our laboratory, the development of our perception of the mechanisms by which the most relevant steps of this pathway occur in bacteria. In particular, we describe: (a) the mechanistics and kinetics of translation initiation; (b) properties of mRNAs with and without Shine-Dalgarno sequence relevant for initiation site selection and translational efficiency; c) ribosomal binding and dissociation of the initiation factors, formation and properties of translation initiation intermediates; (d) the mechanisms by which translation initiation fidelity is ensured. Finally, we provide a short survey of the known translation initiation inhibitors. Copyright © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.