Background: In recent years, medical clowning research has received increasing interest. However, the clowns’ dispositional factors associated with their turnover behavior have yet to be examined systematically.Objectives: The current study examined whether individual differences in humor predict turnover behavior. This was accomplished by investigating the mediating role of the medical clowns’ job satisfaction and the moderating role of previous traumatic experiences.Method: Israeli medical clowns (N = 111) participated in a three-stage longitudinal study. Humorous dispositions were measured during the first week of medical clowning training, job satisfaction was measured two months later, and turnover behaviors were measured after six months.Results: Findings revealed that higher humor appreciation was associated with actual turnover through the mediating role of job satisfaction, whereas humor creation directly increased turnover. In addition, previous traumatic experiences moderated the associations between humor appreciation and turnover.Conclusion: Overall, our research findings support the notion that a humorous disposition can aid in predicting medical clowning turnover. As humor can be observed in numerous settings, in medicine as well as in other fields, we suggest implementing procedures to decrease the turnover rate of medical clowns.