Structural and functional insights into the role of the N-terminal Mps1 TPR domain in the SAC (spindle assembly checkpoint).
The SAC (spindle assembly checkpoint) is a surveillance system that ensures the timely and accurate transmission of the genetic material to offspring. The process implies kinetochore targeting of the mitotic kinases Bub1 (budding uninhibited by benzamidine 1), BubR1 (Bub1 related) and Mps1 (monopolar spindle 1), which is mediated by the N-terminus of each kinase. In the present study we report the 1.8 Å (1 Å=0.1 nm) crystal structure of the TPR (tetratricopeptide repeat) domain in the N-terminal region of human Mps1. The structure reveals an overall high similarity to the TPR motif of the mitotic checkpoint kinases Bub1 and BubR1, and a number of unique features that include the absence of the binding site for the kinetochore structural component KNL1 (kinetochore-null 1; blinkin), and determinants of dimerization. Moreover, we show that a stretch of amino acids at the very N-terminus of Mps1 is required for dimer formation, and that interfering with dimerization results in mislocalization and misregulation of kinase activity. The results of the present study provide an important insight into the molecular details of the mitotic functions of Mps1 including features that dictate substrate selectivity and kinetochore docking.
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Philippe
P
Thebault
Dimitri Y
DY
Chirgadze
Zhen
Z
Dou
Tom L
TL
Blundell
Sabine
S
Elowe
Victor M
VM
Bolanos-Garcia
D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry* / Q000502:physiology*; D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000502:physiology*; D018360:Crystallography, X-Ray; D057809:HEK293 Cells; D006367:HeLa Cells; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry* / Q000502:physiology*; D006801:Humans; D059566:M Phase Cell Cycle Checkpoints / Q000502:physiology*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000502:physiology*; D020449:Repetitive Sequences, Amino Acid
D033942:Adaptor Proteins, Vesicular Transport; D018797:Cell Cycle Proteins; C475144:TTC1 protein, human; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C075840:TTK protein, human; D020962:Heterotrimeric GTP-Binding Proteins