Large river valleys have long been seen as important factors to shape the mobility, communication, and exchange of Pleistocene hunter-gatherers. However, rivers have been debated as either natural entities people adapt and react to or as cultural and meaningful entities people experience and interpret in different ways. Here, we attempt to integrate both perspectives. Building on theoretical work from various disciplines, we discuss the relationship between biophysical river properties and sociocultural river semantics and suggest that understanding a river’s persona is central to evaluating its role in spatial organization. By reviewing the literature and analyzing European Upper Paleolithic site distribution and raw material transfer patterns in relation to river catchments, we show that the role of prominent rivers varies considerably over time. Both ecological and cultural factors are crucial to explaining these patterns. Whereas the Earlier Upper Paleolithic record displays a general tendency toward conceiving rivers as mobility guidelines, the spatial consolidation process after the colonization of the European mainland is paralleled by a trend of conceptualizing river regimes as frontiers, separating archaeological entities, regional groups, or local networks. The Late Upper Paleolithic Magdalenian, however, is characterized again by a role of rivers as mobility and communication vectors. Tracing changing patterns in the role of certain river regimes through time thus contributes to our growing knowledge of human spatial behavior and helps to improve our understanding of dynamic and mutually informed human-environment interactions in the Paleolithic.Electronic supplementary materialThe online version of this article (doi:10.1007/s10816-015-9263-x) contains supplementary material, which is available to authorized users.