title: Activity of the Ste20-like kinase, SLK, is enhanced by homodimerization.
abstract: The expression and activation of the Ste20-like kinase, SLK, is increased during renal development and recovery from ischemic acute renal failure. SLK promotes apoptosis, and during renal injury and repair, transcriptional induction or posttranscriptional control of SLK may, therefore, regulate cell survival. SLK contains protein interaction (coiled-coil) domains, suggesting that posttranslational homodimerization may also modulate SLK activity. We therefore expressed coiled-coil regions in the C-terminal domain of SLK as fusion proteins and demonstrated their homodimerization. By gel-filtration chromatography, endogenous and heterologously expressed SLK were detected in a macromolecular protein complex. To test the role of homodimerization in kinase activation, we constructed a fusion protein consisting of the SLK catalytic domain (amino acids 1-373) and a modified FK506 binding protein, Fv (Fv-SLK 1-373). Addition of AP20187 (an analog of FK506) enhanced the homodimerization of Fv-SLK 1-373. In an in vitro kinase assay, the dimeric Fv-SLK 1-373 displayed greater kinase activity than the monomeric form. In cells expressing Fv-SLK 1-373, homodimerization increased activation-specific phosphorylation of the proapoptotic kinases, c-Jun N-terminal kinase and p38 kinase. Compared with the monomer, dimeric Fv-SLK 1-373 enhanced the activation of a Bax promoter-luciferase reporter. Finally, expression of Fv-SLK 1-373 induced apoptosis, and the effect was increased by homodimerization. Thus the activity, downstream signaling, and functional effects of SLK are enhanced by dimerization of the kinase domain.
authors.affiliation: Department of Medicine, McGill University Health Centre, McGill University, Montreal, Quebec, Canada.
authors.forename: Sierra
authors.initials: S
authors.lastname: Delarosa
authors.forename: Julie
authors.initials: J
authors.lastname: Guillemette
authors.forename: Joan
authors.initials: J
authors.lastname: Papillon
authors.forename: Ying-Shan
authors.initials: YS
authors.lastname: Han
authors.forename: Arnold S
authors.initials: AS
authors.lastname: Kristof
authors.forename: Andrey V
authors.initials: AV
authors.lastname: Cybulsky
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D023421:Models, Animal; D055503:Protein Multimerization / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology; D000099317:Hippo Kinases
chemical_list: D017346:Protein Serine-Threonine Kinases; C402650:SLK protein, mouse; D000099317:Hippo Kinases