Fatigue is a common side effect of tyrosine kinase inhibitor (TKI) therapy in patients with chronic myeloid leukemia (CML). However, the prevalence of TKI-induced fatigue remains uncertain and little is known about predictors of fatigue and its relationship with physical activity. In this study, 220 CML patients receiving TKI therapy and 110 gender- and age-matched controls completed an online questionnaire to assess fatigue severity and fatigue predictors (Part 1). In addition, physical activity levels were objectively assessed for 7 consecutive days in 138 severely fatigued and non-fatigued CML patients using an activity monitor (Part 2). We demonstrated that the prevalence of severe fatigue was 55.5% in CML patients and 10.9% in controls (P<0.001). We identified five predictors of fatigue in our CML population: age (odds ratio [OR] 0.96, 95% confidence interval [95% CI]: 0.93-0.99), female gender (OR 1.76, 95% CI: 0.92-3.34), Charlson Comorbidity Index (OR 1.91, 95% CI: 1.16-3.13), the use of comedication known to cause fatigue (OR 3.43, 95% CI: 1.58-7.44), and physical inactivity (OR of moderately active, vigorously active and very vigorously active compared to inactive 0.43 (95% CI: 0.12-1.52), 0.22 (95% CI: 0.06-0.74), and 0.08 (95% CI: 0.02-0.26), respectively). Objective monitoring of activity patterns confirmed that fatigued CML patients performed less physical activity of both light (P=0.017) and moderate to vigorous intensity (P=0.009). In fact, compared to the non-fatigued patients, fatigued CML patients performed 1 hour less of physical activity per day and took 2,000 fewer steps per day. Our findings facilitate the identification of patients at risk of severe fatigue and highlight the importance of setting reduction of fatigue as a treatment goal in CML care. This study was registered at The Netherlands Trial Registry, NTR7308 (Part 1) and NTR7309 (Part 2).