Temperature sensitivity (Q₁₀) of permafrost carbon (C) release upon thaw is a vital parameter for projecting permafrost C dynamics under climate warming. However, it remains unclear how mineral protection interacts with microbial properties and intrinsic recalcitrance to affect permafrost C fate. Here, we sampled permafrost soils across a 1000-km transect on the Tibetan Plateau and conducted two laboratory incubations over 400- and 28-day durations to explore patterns and drivers of permafrost C release and its temperature response after thaw. We find that mineral protection and microbial properties are two types of crucial predictors of permafrost C dynamics upon thaw. Both high C release and Q₁₀ are associated with weak organo-mineral associations but high microbial abundances and activities, whereas high microbial diversity corresponds to low Q₁₀. The attenuating effects of mineral protection and the dual roles of microbial properties would make the permafrost C-climate feedback more complex than previously thought.