Cue reactivity has great potential to advance our understanding of posttraumatic stress disorder (PTSD), substance use disorder (SUD), and PTSD/SUD comorbidity. The present investigation examined distress tolerance (DT) with regard to trauma and substance cue reactivity. Participants included 58 low-income, inner-city adults (49.1% women; Mage = 45.73, SD = 10.00) with substance dependence and at least 4 symptoms of PTSD. A script-driven cue reactivity paradigm was utilized. Four DT measures were administered, including the Distress Tolerance Scale (DTS), Mirror-Tracing Persistence Task (MTPT), Breath-Holding Task (BH), and Paced Auditory Serial Addition Task (PASAT). Lower DT, as indexed by MTPT duration, was significantly predictive of greater levels of self-reported substance cravings/urges in response to trauma cues, above and beyond covariates. Lower DTS scores predicted lower levels of self-reported control/safety ratings in response to substance cues. None of the DT indices was significantly predictive of heart rate variability. Clinical and research implications are discussed.