title: The molecular basis for phosphodependent substrate targeting and regulation of Plks by the Polo-box domain.
abstract: Polo-like kinases (Plks) perform crucial functions in cell-cycle progression and multiple stages of mitosis. Plks are characterized by a C-terminal noncatalytic region containing two tandem Polo boxes, termed the Polo-box domain (PBD), which has recently been implicated in phosphodependent substrate targeting. We show that the PBDs of human, Xenopus, and yeast Plks all recognize similar phosphoserine/threonine-containing motifs. The 1.9 A X-ray structure of a human Plk1 PBD-phosphopeptide complex shows that the Polo boxes each comprise beta6alpha structures that associate to form a 12-stranded beta sandwich domain. The phosphopeptide binds along a conserved, positively charged cleft located at the edge of the Polo-box interface. Mutations that specifically disrupt phosphodependent interactions abolish cell-cycle-dependent localization and provide compelling phenotypic evidence that PBD-phospholigand binding is necessary for proper mitotic progression. In addition, phosphopeptide binding to the PBD stimulates kinase activity in full-length Plk1, suggesting a conformational switching mechanism for Plk regulation and a dual functionality for the PBD.
authors.affiliation: Center for Cancer Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Andrew E H
authors.initials: AE
authors.lastname: Elia
authors.forename: Peter
authors.initials: P
authors.lastname: Rellos
authors.forename: Lesley F
authors.initials: LF
authors.lastname: Haire
authors.forename: Jerry W
authors.initials: JW
authors.lastname: Chao
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Ivins
authors.forename: Katja
authors.initials: K
authors.lastname: Hoepker
authors.forename: Duaa
authors.initials: D
authors.lastname: Mohammad
authors.forename: Lewis C
authors.initials: LC
authors.lastname: Cantley
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Smerdon
authors.forename: Michael B
authors.initials: MB
authors.lastname: Yaffe
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D018797:Cell Cycle Proteins; D018360:Crystallography, X-Ray; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009154:Mutation; D010748:Phosphopeptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins; D016415:Sequence Alignment; D013379:Substrate Specificity; D014981:Xenopus; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D010748:Phosphopeptides; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D000097869:Polo-Like Kinase 1; C490219:PLK4 protein, human