The yeast Slx5-Slx8 DNA integrity complex displays ubiquitin ligase activity.
Genetic studies in budding yeast have previously implicated SLX5 and SLX8 in the control of genome stability and sumoylation. These genes encode RING-finger domain proteins that form a complex of unknown function. Because RING-finger proteins comprise a large class of ubiquitin (Ub) ligases, Slx5 and Slx8 were tested for this activity. Here we show that the Slx5-Slx8 complex, but not its individual subunits, stimulates several human and yeast Ub conjugating enzymes, including Ubc1, 4, 5, and Ubc13-Mms2. The RING-finger domains of both subunits are genetically required for suppression of slx sgs1Delta synthetic-lethality, and point mutations that abolish Ub ligase activity in vitro also eliminate in vivo complementation. Targets of the in vitro ubiquitination reaction include the Slx5 and Slx8 subunits themselves, and the homologous recombination proteins Rad52 and Rad57. We propose that the Slx5-Slx8 complex functions as a two-component Ub ligase in vivo and that it controls genome stability and sumoylation via ubiquitination.
Ii
Tatsuya
T
Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, New Jersey, USA.
Fung
Jacqueline
J
Mullen
Janet R
JR
Brill
Steven J
SJ
D004271:DNA, Fungal / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D054829:RING Finger Domains / Q000235:genetics / Q000502:physiology; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D043743:Ubiquitin-Protein Ligase Complexes / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D044767:Ubiquitin-Protein Ligases; D016335:Zinc Fingers / Q000502:physiology*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins; D043743:Ubiquitin-Protein Ligase Complexes; C515574:Slx8 protein, S cerevisiae; D044767:Ubiquitin-Protein Ligases; C515573:Slx5 protein, S cerevisiae