Crystal structure of the N-terminal domain of human Timeless and its interaction with Tipin.
Human Timeless is involved in replication fork stabilization, S-phase checkpoint activation and establishment of sister chromatid cohesion. In the cell, Timeless forms a constitutive heterodimeric complex with Tipin. Here we present the 1.85 Å crystal structure of a large N-terminal segment of human Timeless, spanning amino acids 1-463, and we show that this region of human Timeless harbours a partial binding site for Tipin. Furthermore, we identify minimal regions of the two proteins that are required for the formation of a stable Timeless-Tipin complex and provide evidence that the Timeless-Tipin interaction is based on a composite binding interface comprising different domains of Timeless.
Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
Sandro
S
Holzer
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Gianluca
G
Degliesposti
Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
Mairi L
ML
Kilkenny
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Sarah L
SL
Maslen
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK.
Dijana
D
Matak-Vinkovíc
MRC Laboratory of Molecular Biology, Cambridge CB2 0QH, UK.
Mark
M
Skehel
Department of Biochemistry, University of Cambridge, Cambridge CB2 1GA, UK.
Luca
L
Pellegrini
D055592:Biophysical Phenomena; D002352:Carrier Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D003432:Cross-Linking Reagents / Q000378:metabolism; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000378:metabolism*; D013058:Mass Spectrometry; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D055503:Protein Multimerization; D040681:Structural Homology, Protein
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C509769:TIMELESS protein, human; C516227:Tipin protein, human