Toxoplasma gondii progression through the lytic cycle depends on a divergent actin and an array of associated unconventional myosin motors. These acto-myosin systems power mechanical processes, such as active invasion and egress of host cells, cell division and organellar trafficking. How actin-associated proteins regulate the organisation and turnover of the actin filaments to support these diverse processes is poorly understood. Cyclase-associated protein (CAP) is an actin-binding protein conserved across eukaryotes. We show that Toxoplasma CAP has a unique extension that is not present in most other Apicomplexa, giving rise to two isoforms with distinct subcellular localisations: one is localised at the parasite apex while the other is cytosolic. Here we investigated the role of CAP in Toxoplasma biology using a new generation of RH ∆ku80∆hxgprt DiCre parasites where loss of DiCre activity is prevented. Conditional knockout of CAP led to significant defects in motility, invasion, active egress, dense granule trafficking, daughter cell orientation, juxtanuclear accumulation of actin and cell-cell communication but only modest defects in synchronicity of division and no defect in replication of the apicoplast. Despite displaying phenotypes closely resembling the actin knockout, CAP is dispensable for in vitro culture. Strikingly, CAP knockout in the type I RH parasite strain does not affect in vivo virulence but results in complete attenuation of the type II Pru strain. 3D electron microscopy reveals that loss of CAP results in a defect in formation of a normal central residual body, but parasites remain connected within the vacuole. This dissociates synchronicity of division and parasite rosetting and reveals that establishment and maintenance of the residual body may be more complex than previously thought. These results highlight the different spatial requirements for F-actin dynamics in Toxoplasma that depend, in part, on CAP function.