Recent work suggests that Drosophila and Saccharomyces yeasts may establish a mutualistic association, and that this is driven by chemical communication. While individual volatiles have been implicated in the attraction of D. melanogaster, the semiochemicals affecting the behavior of the sibling species D. simulans are less well characterized. Here, we scrutinized a broad range of volatiles produced by attractive and repulsive yeasts to experimentally evaluate the chemical nature of communication between these species. When grown in liquid or on agar-solidified grape juice, attraction to S. cerevisiae was driven primarily by 3-methylbutyl acetate (isoamyl acetate) and repulsion by acetic acid, a known attractant to D. melanogaster (also known as vinegar fly). By using T-maze choice tests and synthetic compounds, we showed that these responses are strongly influenced by compound concentration. Moreover, the behavioral response is impacted further by the chemical context of the environment. Thus, chemical communication between yeasts and flies is complex, and is not driven simply by the presence of single volatiles, but modulated by compound interactions. The ecological context of chemical communication needs to be taken into consideration when testing for ecologically realistic responses.