Tuberculosis is a chronic bacterial infection caused by Mycobacterium tuberculosis, which persists in the host despite immune responses. This study shows that M. tuberculosis uses host cholesterol as a carbon and energy source, particularly during chronic infection. The mce4 gene cluster encodes a cholesterol import system essential for bacterial survival in the lungs and in IFN-γ-activated macrophages. Cholesterol import is not required for initial infection or growth in resting macrophages but is critical for persistence. The ability to catabolize cholesterol allows M. tuberculosis to survive in nutrient-poor environments created by IFN-γ, which sequesters pathogens in compartments lacking common nutrients. The study demonstrates that M. tuberculosis can degrade cholesterol, using it for energy and carbon, and that the Mce4 transporter is crucial for this process. The loss of this function impairs bacterial growth during chronic infection and in IFN-γ-activated macrophages. The findings suggest that cholesterol utilization is a key strategy for M. tuberculosis to maintain chronic infection by accessing host cholesterol in macrophages. The study also highlights the importance of cholesterol in the intracellular environment of macrophages and its role in bacterial survival. The results indicate that Mce4 is a major cholesterol import system in M. tuberculosis, essential for its survival and persistence in chronic infections.Tuberculosis is a chronic bacterial infection caused by Mycobacterium tuberculosis, which persists in the host despite immune responses. This study shows that M. tuberculosis uses host cholesterol as a carbon and energy source, particularly during chronic infection. The mce4 gene cluster encodes a cholesterol import system essential for bacterial survival in the lungs and in IFN-γ-activated macrophages. Cholesterol import is not required for initial infection or growth in resting macrophages but is critical for persistence. The ability to catabolize cholesterol allows M. tuberculosis to survive in nutrient-poor environments created by IFN-γ, which sequesters pathogens in compartments lacking common nutrients. The study demonstrates that M. tuberculosis can degrade cholesterol, using it for energy and carbon, and that the Mce4 transporter is crucial for this process. The loss of this function impairs bacterial growth during chronic infection and in IFN-γ-activated macrophages. The findings suggest that cholesterol utilization is a key strategy for M. tuberculosis to maintain chronic infection by accessing host cholesterol in macrophages. The study also highlights the importance of cholesterol in the intracellular environment of macrophages and its role in bacterial survival. The results indicate that Mce4 is a major cholesterol import system in M. tuberculosis, essential for its survival and persistence in chronic infections.