Coatomer, the cytosolic protein complex, consists of seven subunits (alpha-, beta-, beta'-, gamma-, delta-, epsilon-, and zeta-COP) and is involved in vesicle trafficking early in the secretory pathway in collaboration with a 20 kDa GTP-binding protein (ARF). In the present study, we have identified a yeast gene which encodes a protein having 39% amino acid sequence identity with bovine zeta-COP. This gene (YZC1 for Yeast Zeta COP) is essential for vegetative growth and the growth defect of delta yzc1 cells was restored by bovine zeta-COP cDNA. We isolated a temperature-sensitive mutant of YZC1 (yzc1ts) and examined its capacity for both the ER-to-Golgi transport and the double lysine motif (KKXX)-mediated retrograde transport from Golgi to ER. At non-permissive temperature, the yzc1ts cells exhibited a weak defect in the anterograde transport, but a strong defect in the retrograde vesicle transport. We conclude that Yzc1p is a yeast homologue of mammalian zeta-COP and participates mainly in the Golgi-to-ER retrograde transport.