Streptococcus pneumoniae (the pneumococcus) is a major global cause of human disease. Since the publication of the entire sequence of TIGR4 in 2001, our understanding of this human pathogen has increased significantly. Genetic studies, and the use of mutant strains have refined our understanding of the pathogenic mechanisms of classic pneumococcal virulence factors, including the polysaccharide capsule, pneumolysin and surface-expressed proteins. Genetic screens are identifying novel virulence factors. Characterization of pili and bacteriocins, as well as genes associated with competence, metabolism and resistance to oxidative stress has provided new insights into the genetic diversity of the pneumococcus. Further appreciation of the molecular basis of pneumococcal pathogenesis will lead to more effective strategies for the prevention and management of pneumococcal disease.