Type B trichothecenes, deoxynivalenol (DON) and nivalenol (NIV), are secondary metabolites of Fusarium species and are major pollutants in food and feed products. Recently, the production trend of their derivatives, 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON), and 4-acetylnivalenol (4-AcNIV or fusarenon-X), has been changing in various regions worldwide. Although in vivo behavior has been reported, it is necessary to acquire more detailed information about these derivatives. Here, the yeast PDR5 mutant was used for toxicity evaluation, and the growth test revealed that DON, 15-AcDON, and 4-AcNIV had higher toxicity compared to 3-AcDON and NIV. 15-AcDON exerted the most significant gene expression changes, and cellular localization clustering exhibited repression of mitochondrial ribosomal genes. This study suggests that the toxicity trends of both DON products (DON and its derivatives) and NIV products (NIV and its derivatives) are similar to those observed in mammalian cells, with a notable toxic response to 15-AcDON.