Some studies have suggested that non‐native species invasions may threaten local diversity by creating homogenized environments. However, many studies have been based on limited or anecdotal data, and/or have failed to consider the influence of habitat modification together with possible influences of non‐native species on native ones. Hemidactylus mabouia (Squamata, Gekkonidae) likely invaded natural environments in Brazil hundreds of years ago. Yet, little is known about whether it affects native lizard fauna. We tested whether H. mabouia negatively influences native lizard species richness and abundance on a regional scale and locally through niche overlap. We analyzed species abundance and richness of nine lizard assemblages, in five of which H. mabouia occurred. We evaluated niche overlap of species in a lizard assemblage with high H. mabouia abundance through null models. Niche axes included spatial use, temporal activity and diet. Although species abundance did not differ among sites with and without the invasive species, the presence of H. mabouia seems constrained to the richer assemblages sampled. We observed significantly higher niche overlap in spatial (ɸobs = 0.63; ɸexp = 0.37; Pobs ≥ Pexp = 0.0002) and trophic axes (ɸobs = 0.46; ɸexp = 0.17; Pobs ≥ Pexp < 0.001), but not in activity. When we considered all axes (three‐dimensional niche), there was no overlapping among the lizard species. Our findings did not support the hypothesis that this non‐native species negatively influences other sympatric lizard species.