The transmembrane domains (TMDs) of many type I integral membrane proteins contain determinants that cause localization in the endoplasmic reticulum (ER) in mammalian cells by an unknown mechanism. Here we show that the yeast ER localization machinery recognizes determinants in TMDs that are very similar to those identified previously in mammalian cells. These determinants are recognized in post-ER compartments and recycled back to the ER, thus acting as ER retrieval signals. Moreover determinants in TMDs are inefficiently sorted in several previously characterized yeast mutants with defects in the ER retrieval machinery. Similar ER retrieval signals are also recognized in the TMDs of polytopic integral membrane proteins, apparently by the same sorting machinery. The isolation of new mutants defective in sorting of membrane determinants might provide a better understanding of the molecular mechanisms involved in this process.