Purpose The purpose of this paper is to test the mediating role of autonomy in the effect of a pro-innovation organizational climate and supervisor supportiveness (SS) on the innovative behavior (IB) of nurses. Design/methodology/approach The sample of this cross-sectional, correlational study consisted of 332 nurses, who were selected, on the basis of the convenience sampling method, from among the nurses working at two public university hospitals in Istanbul. The data of the study were collected between February and May of 2015. Descriptive analysis and linear regression analysis were used for data analysis, and the bootstrapping method was applied to test the significance of the mediating role. Findings The model used for examining the mediating role of autonomy was found to be statistically significant, as it explained 36 percent of the variance of IB. When the significance of the mediating role was tested, its effect on both innovation climate and SS was observed to be significant. Originality/value The obtained results indicated that a pro-innovation organizational climate and SS functioned as premises for autonomy. In turn, the rise in autonomy level that served to innovative climate and SS increased the IB of the nurses through boosting their autonomy level. From these results, it can be argued that nurses with a high level of professional autonomy can create innovative outputs and provide added value through the IB they practice in providing patient care and general health services.