Media literacy is a critical component for digital citizenship; however, the nature of this relationship and the role of other mediating variables such as digital literacy in this relationship are not empirically tested in various contexts. This paper examined the effect of media literacy on digital citizenship and tested the mediating role of digital literacy in this relationship. Mediation model was applied to analyze the relationships between the variables, based on data from 555 pre-service teachers in Turkey. This study revealed a direct relationship between media literacy and digital citizenship. In addition, digital literacy partially mediated the effect of pre-service teachers' media literacy levels on their digital citizenship. Results are discussed and implications for practitioners and recommendations for future research are offered.