Mus81, a fission yeast protein related to the XPF subunit of ERCC1-XPF nucl
eotide excision repair endonuclease, is essential for meiosis and important
for coping with stalled replication forks. These processes require resolut
ion of X-shaped DNA structures known as Holliday junctions. We report that
Mus81 and an associated protein Eme1 are components of an endonuclease that
resolves Holliday junctions into linear duplex products. Mus81 and Eme1 ar
e required during meiosis at a late step of meiotic recombination. The mus8
1 meiotic defect is rescued by expression of a bacterial Holliday junction
resolvase. These findings constitute strong evidence that Mus81 and Emel1 a
re subunits of a nuclear Holliday junction resolvase.