Staphylococcus aureus and most other Gram-positive bacteria contain characteristic membrane- or peptidoglycan-attached carbohydrate-based polymers in their cell walls whose structure, function, and biosynthesis is only superficially understood. The composition of these cell wall glycopolymers (CWG) is highly variable and often species and strain specific. CWGs have been implicated in adherence to host cells, activation of immune responses e.g. via the toll-like receptors (TLRs) and susceptibility to antibiotics and antimicrobial host defenses. Accordingly, the peptidoglycan-anchored wall teichoic acid (WTA) plays a crucial role in S. aureus nasal colonization, in endovascular infections, and in biofilm formation. CWGs are also thought to be important in bacterial physiology by controlling autolytic enzymes, storing divalent cations, or attaching surface proteins. Recent studies have yielded a growing picture of the biosynthetic pathways of WTA and the membrane anchored lipoteichoic acid (LTA). Accordingly, CWG represent a very important topic for research that may help to identify candidate targets for new antimicrobials, vaccines, and diagnostics.