title: The small GTPase Rac interacts with ubiquitination complex proteins Cullin-1 and CDC23.
abstract: Racs are involved in the regulation of important cellular processes including mitogenesis. We found that the E3 ubiquitination ligase subunit Cullin-1 interacts with constitutively active Rac3 but not with wild-type Rac3 in yeast. In mammalian cell lysates, Cullin-1 bound to V12Rac3, effector domain mutants V12L37Rac3 and V12H40Rac3, and insert domain deletion mutant V12Rac3DeltaIns(124-135). Cullin-1 also formed a clearly detectable complex with other activated Rac3-related proteins including Rac1, Rac2, Cdc42 and RhoA but not with the distantly related small GTPase Rap1. Since the proteasome is involved in cell cycle control through the programmed degradation of cell cycle proteins, the possible regulation of Rac levels during the cell cycle was examined. However, Rac was expressed at constant levels throughout the cell cycle, and a specific proteasome inhibitor had no effect on Rac protein levels. These combined results indicate that the binding of activated Rac to Cullin-1 does not affect Rac protein levels, nor does it mediate the regulation of mitogenesis by Rac. However, Rac-Cullin-1 interactions may serve to regulate other E3 ligase functions such as subcellular localization. Indeed, activated Rac3 and Cullin-1 co-localized to the perinuclear region of the cell. We also detected complex formation between Rac and the APC component CDC23. These results indicate that Rac may regulate specific proteolytic processes through directed subcellular localization of SCF or APC complexes.
authors.affiliation: Division of Hematology/Oncology, Section of Molecular Carcinogenesis, Childrens Hospital of Los Angeles Research Institute, Los Angeles, CA 90027, USA.
authors.forename: D
authors.initials: D
authors.lastname: Senadheera
authors.forename: L
authors.initials: L
authors.lastname: Haataja
authors.forename: J
authors.initials: J
authors.lastname: Groffen
authors.forename: N
authors.initials: N
authors.lastname: Heisterkamp
mesh_terms: D000818:Animals; D064196:Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome; D016466:CHO Cells; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D006224:Cricetinae; D044842:Cullin Proteins; D006801:Humans; D025801:Ubiquitin; D020744:rac GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D064196:Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome; C577440:CDC23 protein, human; D018797:Cell Cycle Proteins; C100318:Cullin 1; D044842:Cullin Proteins; C411563:RAC3 protein, human; D025801:Ubiquitin; D020744:rac GTP-Binding Proteins