Abundance, affordability, and also the usability of sulfur as a part of the binder are the main reasons for using this additive in asphalt mixtures. However, lack of proper adhesion and brittle behavior of sulfur-modified asphalt mixtures could make them susceptible to moisture damage and fatigue cracking. In this regard, anti-stripping agents and some polymers like crumb rubber may be used to overcome these types of distresses. This research aimed at investigating the effects of polymeric sulfur and crumb rubber on the moisture sensitivity and fatigue behavior of asphalt mixtures. To this end, asphalt mixtures were subjected to 1 and 3 freeze-thaw cycles. Then, an indirect tensile fatigue test was carried out. Moreover, the response surface method (RSM) was used to assess the interaction between various parameters in samples containing polymeric sulfur and crumb rubber. Results showed that with an increase in polymeric sulfur content, the fatigue life was reduced and the moisture sensitivity was increased. Additionally, the RSM was found to be effective in ranking parameters influencing the performance of asphalt mixtures. According to the experimental results, a crumb rubber-modified binder could improve the fatigue life of the polymeric sulfur-modified mixture up to 70%. Furthermore, experimental results and RSM analysis indicated that crumb rubber would be more effective in higher numbers of freeze-thaw cycles and raised temperatures, in which polymeric sulfur-modified asphalt mixtures are probably prone to moisture damages (i.e. the lack of adequate cohesion and adhesion).