This paper presents the findings of a study on the impact of the transformation of a road transport corridor on the form of a functio-spatial structure, as determined by the placement of significant economic activity sites within this corridor. The investigation of relations and interdependencies in the development of the road transport system and development structure transformation processes allowed for the identification of tendencies in the shaping of space, as well as for building models that reflect the transformation of the road corridor under study. The study focused on a fragment of the road transport corridor between Kraków and Bochnia (called the “Kraków East–Bochnia” corridor), as a distinctive case of contemporary transformations of the functio-spatial structure that happen under the influence of the construction and opening of a section of highway A4. The study was based on original field work, a review of the literature and an analysis of applicable planning documents. GIS tools, cartographic resources and satellite images were also used. The transformation of the area under investigation and the increase in its accessibility due to the presence of the highway pointed to the area’s high attractiveness in terms of real estate development—especially at nodal sites along the linkages that connected the highway with other elements of the road corridor. The relationships between the completed highway section and the previous road layout are crucial to the emergence of economic activity sites and areas, and thus lead to a transformation of developed space following a new spatial model.