Purpose This study sought to investigate the effects of role conflict, role ambiguity, and workload on burnout syndrome among clinical nurses working in university hospitals and the mediating role of work-related stress in these effects. Design and Methods Cross-sectional questionnaire-based survey was conducted. Data from 386 nurses were evaluated. The research variables were analyzed using statistical methods such as factor analysis, correlation analysis, regression analysis, Sobel tests, and bootstrapping method. Findings It was determined that role conflict, role ambiguity, workload, and work-related stress affect nurses' burnout significantly and work-related stress has a mediator role between burnout and the independent variables. Practice Implications Nurses' burnout syndrome can be prevented if managerial practices to reduce the sources of role-related stressors and work-related stress are implemented by managers.