Ligation of the ubiquitin-like protein SUMO (Smt3p) to other proteins is es
sential for viability of the yeast Saccharomyces cerevisiae. Like ubiquitin
(Ub), SUMO undergoes ATP-dependent activation by a specific activating enz
yme. SUMO-activating enzyme is a heterodimer composed of Uba2p and Aos1p, p
olypeptides with sequence similarities, respectively, to the C- and N-termi
nal parts of Ub-activating enzyme. To study the function of SUMO conjugatio
n, we isolated uba2 mutants that were temperature-sensitive for growth. In
these mutants conjugation of SUMO to other proteins was drastically reduced
, even at the temperature permissive for growth. In a screen for spontaneou
s suppressors of the temperature-sensitive growth phenotype of the mutant u
ba2-ts9, we isolated a strain with a null mutation (sut9) in a gene of hith
erto unknown function (SUT9/YIL031W/SMT4). This gene encodes a protein with
similarities to Ulp1p, a dual-function protease that processes the SUMO pr
ecursor and deconjugates SUMO from its substrates. The novel protein was th
erefore termed Ulp2p. Inactivation of ULP2 in a strain expressing wild-type
SUMO-activating enzyme resulted in slow and temperature-sensitive growth,
and accumulation of SUMO conjugates. Thus, mutations in SUMO-activating enz
yme and mutations in Ulp2p suppress each other, indicating that SUMO conjug
ation and deconjugation must be in balance for cells to grow normally. Othe
r phenotypes of ulp2 mutants include a defect in cell cycle progression, hy
persensitivity to DNA damage, and chromosome mis-segregation. Ulp2p is pred
ominantly located within the nucleus, whereas Ulp1p colocalizes with nuclea
r pore complex proteins, indicating that the apparently distinct functions
of the two SUMO deconjugating enzymes are spatially separated.