Abstract The puma Puma concolor is the fourth largest wild felid and the most widespread native terrestrial mammal of the Americas. We synthesised published literature documenting the biotic interactions of pumas, in order to: 1) advance our understanding of the ecological roles pumas play in natural systems, and 2) support strategic decision‐making about conservation investments, public education, and whole‐ecosystem conservation management. We divided puma biotic interactions into five categories: 1) diet and prey regulation, 2) fear effects on prey (including trophic cascades), 3) effects via carrion production, 4) effects on other carnivores, and 5) ecosystem services. We reviewed 162 studies that met our search criteria, which described 543 ecological interactions between pumas and 485 other species. Puma diet and prey regulation was the most common research topic. The geographic distribution of research was highly skewed towards the USA and Canada, and research in Tropical moist forests was underrepresented. We found a steep increase in the number of scientific publications exploring the biotic interactions associated with pumas over time, but publications that reported effect sizes or measured the strength of interactions did not increase as quickly. We noted numerous gaps in our knowledge of puma biotic interactions and found few well‐controlled studies of prey fear effects, trophic cascades, or ecosystem services. We conclude that pumas are influential ecological actors in natural systems and important brokers of energy and nutrients throughout ecosystems in the Western Hemisphere, linking disparate species from many trophic levels. Ultimately, we found evidence for investing in and prioritising the protection and restoration of puma populations to conserve biodiversity in the Americas.