The present study examines future teachers' motivations for teaching using the FIT-Choice (Factors Influencing Teaching Choice) scale. The focus thereby is on subject interest, a factor that has rarely been accounted for by FIT-Choice studies although it is considered one of the most important factors to students for choosing teaching as a career. It is also assumed that students of different subject domains belong to different subcultures and therefore differ in their motivations. On the basis of n = 386 first-year, Bachelor students qualifying for lower and upper secondary schools from a large university in Germany, a latent confirmatory factor analysis shows that the FIT-Choice scale structure could be replicated and subject-specific interest was rated the most important factor by pre-service teachers. Latent path analyses reveal that students from different subject domains differ slightly in their motivations. More importantly, students who value their studied subjects' importance highly also show higher intrinsic, social-altruistic, and pedagogical motivations.