We studied the effects of environmental driving factors (maximum depth, visibility, oxygen, temperature, and prey densities) on the distribution and diet composition of Nile perch (Lates niloticus) in south-east Lake Victoria from 2009 to 2011. We tested the hypotheses that (i) Nile perch distribution is regulated by the same environmental factors on a local scale (Mwanza Gulf) and on a regional scale (Mwanza Gulf, Speke Gulf and the open lake in Sengerema district), and (ii) driving factors act differently on different Nile perch size classes. Fish were sampled with gillnets. Nile perch densities were highest in the shallow part of the Mwanza Gulf and during the wet seasons, mainly caused by high densities of juveniles. The environmental driving factors explained Nile perch distributions on both regional and local scales in a similar way, often showing non-linear relationships. Maximum depth and temperature were the best predictors of Nile perch densities. Prey densities of shrimp and haplochromines did not strongly affect Nile perch distributions, but did explain Nile perch diet on a local and regional scale. We conclude that abiotic variables drive Nile perch distributions more strongly than prey densities and that feeding takes place opportunistically.