title: The mixed lineage kinase SPRK phosphorylates and activates the stress-activated protein kinase activator, SEK-1.
abstract: SPRK (also called PTK-1 and MLK-3), a member of the mixed lineage kinase subfamily of (Ser/Thr) protein kinases, encodes an amino-terminal SH3 domain followed by a kinase catalytic domain, two leucine zippers interrupted by a short spacer, a Rac/Cdc42 binding domain, and a long carboxyl-terminal proline-rich region. We report herein that SPRK activates the stress-activated protein kinases (SAPKs) but not ERK-1 during transient expression in COS cells; the p38 kinase is activated modestly (1.3-2 fold) but consistently. SPRK also activates cotransfected SEK-1/MKK-4, a dual specificity kinase which phosphorylates and activates SAPK. Reciprocally, expression of mutant, inactive SEK-1 inhibits completely the basal and SPRK-activated SAPK activity. Immunoprecipitated recombinant SPRK is able to phosphorylate and activate recombinant SEK-1 in vitro to an extent comparable to that achieved by MEK kinase-1. These results identify SPRK as a candidate upstream activator of the stress-activated protein kinases, acting through the phosphorylation and activation of SEK-1.
authors.affiliation: Diabetes Unit and Medical Service, Massachusetts General Hospital, Boston, Massachusetts 02129, USA.
authors.forename: A
authors.initials: A
authors.lastname: Rana
authors.forename: K
authors.initials: K
authors.lastname: Gallo
authors.forename: P
authors.initials: P
authors.lastname: Godowski
authors.forename: S
authors.initials: S
authors.lastname: Hirai
authors.forename: S
authors.initials: S
authors.lastname: Ohno
authors.forename: L
authors.initials: L
authors.lastname: Zon
authors.forename: J M
authors.initials: JM
authors.lastname: Kyriakis
authors.forename: J
authors.initials: J
authors.lastname: Avruch
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002460:Cell Line; D018076:DNA, Complementary; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D051381:Rats
chemical_list: D018076:DNA, Complementary; C490205:Sprk protein, rat; D017346:Protein Serine-Threonine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases