Drought stress is a major limiting factor during the process of plant growth and development, especially in arid and semi-arid regions. MYB transcription factors play vital roles in the regulation of many developmental processes under various stresses. The aim of this study was to determine whether PtrMYB119 enhanced dehydration tolerance in Nicotiana tabacum. PtrMYB119, with a weak transactivation activity, was distributed throughout the cell with no apparent specificity. The transgenic tobacco overexpressing PtrMYB119 might regulate dehydration tolerance through increased ABA content and antioxidant enzyme activities, decreased MDA levels, and up-regulation of antioxidant genes, polyamine biosynthesis genes, and drought-responsive genes. Overall, our results could contribute to the elucidation of drought tolerance underlying PtrMYB119 action in tobacco and indicated that PtrMYB119 could be exploited for engineering drought-enduring plants in the future.