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The dynamics of fish communities at tropical and sub-tropical rocky reefs are influenced in many cases by predation activity and predator-prey interactions. These processes usually follow specific diel patterns in reef areas with higher rates of these interactions occurring during the crepuscular periods. However, other factors such as habitat complexity and species-specific behavior may alter these patterns, increasing variability in species interactions. A better understanding of the dynamics of these patterns and processes would allow us to manage and monitor fish communities in these productive and vulnerable areas more efficiently. We investigated behavioral changes of predators and prey fish in sub-tropical "live-bottom" (sandstone) reefs at Gray's Reef National Marine Sanctuary (GRNMS), located 20 nautical miles off the coast of Georgia, USA, using fisheries acoustic methods in association with visual census and direct observation using SCUBA. Changes in co-location and habitat preferences of predators and prey over time throughout the diel cycle were investigated using species distribution models (MAXENT) based on habitat predictors and by means of spatial statistics. The results indicate that predator and prey distribution patterns changed considerably throughout the day. Prey and predator species exhibited complex spatial dynamics and behavior over diel periods, with prey modifying patterns of habitat use and spatial distribution, likely as a response of their interactions with predators. Crepuscular periods were confirmed to be the most active phases in terms of predator-prey interactions and consequently the most variable. The combination of tools and approaches used in this study provided valuable sources of information that support the inferences of predation risk-driven habitat selection of prey in this sub-tropical reef system., Competing Interests: The authors have declared that no competing interests exist.