ObjectiveTo examine the association of perceived distress tolerance—one’s ability to endure aversive experiences—with subsequent substance use frequency during adolescence. MethodsHigh school students (N = 3,203) were surveyed semiannually from 2013 to 2017. The Distress Tolerance Scale (DTS) was administered at baseline (9th grade) and self-reported past 30-day number of days used of alcohol, cigarettes, e-cigarettes, cannabis, and opioids (range: 0–30) were assessed at all timepoints. Self-reported baseline ever (vs. never) use of each substance and depressive/anxiety symptom status (clinical and subclinical vs. none) were tested as moderators. ResultsAmong baseline never substance users, a greater baseline mean DTS score was associated with fewer days of alcohol, cigarette, and opioid use in past-30 days over 4 years of follow-up (IRR range = 0.60–0.84); among ever-users, baseline mean DTS was not significantly associated with substance use across follow-up timepoints (p-interaction = 0.03). The association of DTS and frequency of substance use did not differ by baseline mental health status for any substance (all ps > 0.05). ConclusionsGreater perceived distress tolerance at the start of high school may be a protective factor against substance use frequency prior to initial use. Interventions that build distress tolerance in early adolescence may be warranted to prevent higher frequency substance use throughout high school.