1 / 12

The “deep” characterization of Aurora A N-terminus

The “deep” characterization of Aurora A N-terminus. Isabel Peset Martin. Vanessa Ferreira Leire Mendizabal Oiarzabal. Substrate. Cell localization. Effect of substrates-Aurora interaction. Aur A. PP1, p53, Cdh-1, TPX-2, RasGAP, Ajuba,TACC. Mitotic spindle; centrosome.

deo
Download Presentation

The “deep” characterization of Aurora A N-terminus

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. The “deep” characterization of Aurora A N-terminus IsabelPeset Martin. Vanessa Ferreira Leire Mendizabal Oiarzabal

  2. Substrate Cell localization Effect of substrates-Aurorainteraction Aur A PP1, p53, Cdh-1, TPX-2, RasGAP, Ajuba,TACC Mitotic spindle; centrosome Spindle assembly; cytokinesis; centrosome maturation and separation Aur B Histone H3; INCENP; CENP-A; desmin; Rec-8; vimentin; MCAK; survivin Centrosome; central spindles; chromosome arms Chromosome alignment and segregation; cytokinesis; microtubule dynamics Aur C Aurora B; INCENP Central spindles; chromosome arms? Role in spermatogenesis; possible role in regulation of chromosome segregation and cytokinesis Human Aurora kinases A–C exhibit differential substrate affinity, subcell localization and associated activities Paul D Andrews Oncogene (2005)

  3. DOMAIN STRUCTURE OF Aurora kinases A–C SourceDomainStartEnd PfamA PKinase 133 383 SMART STKc(hidden) 133 383 AURORA A SourceDomainStart End PfamB Pfam-B 34136 1 62 SMART STKc(hidden) 77 327 PfamA PKinase 77 327 seg Low Complexity (hidden) 204 215 AURORA B SourceDomainStartEnd PfamA PKinase 43 293 SMART STKc(hidden) 43 293 AURORA C

  4. Amino acid sequence alignement of human Aurora kinases A–C. aurB 1 MAQKENSYPWPYGRQTAPSG-------------------LSTLPQRVLR----KEPVTPSaurC 1 ----------------------------------------MSSPRAVVQ----LGKAQPAaurA 1 MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVLCPSNSSQRVPLQaurB 38 ALVLMSRSN---------VQPTAAPG------------------------QKVMENSSGTaurC 17 GEELATAN-----------QTAQQPS------------------------SPAM------aurA 61 AQKLVSSHKPVQNQKQKQLQATSVPHPVSRPLNNTQKSKQPLPSAPENNPEEELASKQKNaurB 65 PDILTRHFTIDDFEIGRPLGKGKFGNVYLAREKKSHFIVALKVLFKSQIEKEGVEHQLRRaurC 36 -----RRLTVDDFEIGRPLGKGKFGNVYLARLKESHFIVALKVLFKSQIEKEGLEHQLRRaurA 121 EESKKRQWALEDFEIGRPLGKGKFGNVYLAREKQSKFILALKVLFKAQLEKAGVEHQLRRaurB 125 EIEIQAHLHHPNILRLYNYFYDRRRIYLILEYAPRGELYKELQKSCTFDEQRTATIMEELaurC 91 EIEIQAHLQHPNILRLYNYFHDARRVYLILEYAPRGELYKELQKSEKLDEQRTATIIEELaurA 181 EVEIQSHLRHPNILRLYGYFHDATRVYLILEYAPLGTVYRELQKLSKFDEQRTATYITELaurB 185 ADALMYCHGKKVIHRDIKPENLLLGLKGELKIADFGWSVHAPSLRRKTMCGTLDYLPPEMaurC 151 ADALTYCHDKKVIHRDIKPENLLLGFRGEVKIADFGWSVHTPSLRRKTMCGTLDYLPPEMaurA 241 ANALSYCHSKRVIHRDIKPENLLLGSAGELKIADFGWSVHAPSSRRTTLCGTLDYLPPEMaurB 245 IEGRMHNEKVDLWCIGVLCYELLVGNPPFESASHNETYRRIVKVDLKFPASVPTGAQDLIaurC 211 IEGRTYDEKVDLWCIGVLCYELLVGYPPFESASHSETYRRILKVDVRFPLSMPLGARDLIaurA 301 IEGRMHDEKVDLWSLGVLCYEFLVGKPPFEANTYQETYKRISRVEFTFPDFVTEGARDLIaurB 305 SKLLRHNPSERLPLAQVSAHPWVRANSRRVLPPSALQSVA---aurC 271 SRLLRYQPLERLPLAQILKHPWVQAHSRRVLPPCAQMAS----aurA 361 SRLLKHNPSQRPMLREVLEHPWITANSSKPSNCQNKESASKQS

  5. human 208 LILEYAPLGTVYRELQK--LSKFDEQRTATYITELANALSYCHSKRVIHRDIKPENLLLGmus 199 LILEYAPLGTVYRELQK--LSKFDEQRTATYITELANALSYCHSKRVIHRDIKPENLLLGxenla 215 LILDYAPGGELFRELQK--CTRFDDQRSAMYIKQLAEALLYCHSKKVIHRDIKPENLLLGzebrafish 154 LILEYAPRGEMYKELQR--YGHFDDQRTATYMEEVSDALQYCHEKKVIHRDIKPENLLLGdrome 239 LILEYAPQGTLFNALQAQPMKRFDERQSATYIQALCSALLYLHERDIIHRDIKPENLLLGworm 119 VILDYASRGELFNVLQSQPGHKVNEVIAGRFVRQLANALHYCHSKGVIHRDIKPENLLLDyeast 179 LLMEYLVNGEMYKLLRL--HGPFNDILASDYIYQIANALDYMHKKNIIHRDIKPENILIGhuman 266 SAGELKIADFGWSVHAP-SSRRTTLCGTLDYLPPEMIEGRMHDEKVDLWSLGVLCYEFLVmus 257 SNGELKIADFGWSVHAP-SSRRTTMCGTLDYLPPEMIEGRMHDEKVDLWSLGVLCYEFLVxenla 273 SNGELKIADFGWSVHAP-SSRRTTLCGTLDYLPPEMIEGRMHDETVDLWSLGVLCYEFLVzebrafish 212 YRGELKIADFGWSVHAP-SLRRRTMCGTLDYLPPEMIEGHSHDEKVDLWSIGVLCYECLVdrome 299 HKGVLKIADFGWSVHEP-NSMRMTLCGTVDYLPPEMVQGKPHTKNVDLWSLGVLCFELLVworm 179 SKLNLKLADFGWSVVAD-HSKRHTLCGTMDYLAPEMVSNQPHDFNVDIWAIGILLFEMLVyeast 237 FNNVIKLTDFGWSIINPPENRRKTVCGTIDYLSPEMVESREYDHTIDAWALGVLAFELLThuman 325 GKPPFEANTYQETYKRISRVEFTFPDFVTEGARDLISRLLKHNPSQRPMLREVLEHPWITmus 316 GMPPFEAHTYQETYRRISRVEFTFPDFVTEGARDLISRLLKHNASQRLTLAEVLEHPWIKxenla 332 GKPPFETDTHQETYRRISKVEFQYPPYVSEEARDLVSKLLKHNPNHRLPLKGVLEHPWIIzebrafish 271 GNPPFETRQHAETYKRITKVDLQFPKLVSEGARDLISKLLRHSPSMRLPLRSVMEHRGVKdrome 358 GHAPFYSKNYDETYKKILKVDYKLPEHISKAASHLISKLLVLNPQHRLPLDQVMVHPWILworm 238 GYAPFANQTGDKLIARIKECKIYIPSVVTDGAASLINAIIKKEPQERLPLVDIMAHPWIKyeast 297 GAPPFEEEMKDTTYKRIAALDIKMPSNISQDAQDLILKLLKYDPKDRMRLGDVKMHPWILhuman 385 ANSSKP-SNCQNKESASKQS---------mus 376 ANSSKPPTGHTSKEPTSKSS---------xenla 392 KNSQLK----KKDEPLPGAQ---------zebrafish 331 ANSRRVLPPVCSSDPH-------------drome 418 AHTQ-------------------------worm 298 EMKQREDIEVPLFISTLTKSSRNNSTANQyeast 357 RNKPFWENKRL------------------ human 1 ------MDRS-KENCISGPVKATAPVGG-PKRVLVTQQFPCQNPLP------VNSGQAQRmus 1 ------MDRC-KENCVSRPVKTTVPFG--PKRVLVTEQIPSQNLGS------ASSGQAQRxenla 1 ------MERAVKENHKPSNVKIFHPMTEGAKRIPVNQPQSTQFRPP------GTAVSAQRzebrafish 1 --------------------------------VFAVDREAEEKKHT------RTGGTLD-drome 1 MSHPSDHVLRPKENAPHRMPEKSAAVLNMQKNLLLGKKPNSENMAPDSKPLPGSSGALIRworm 1 -------------------------------------MSGKENTAP--------------yeast 1 MQRNSLVNIKLNANSPSKKTTTRPNTSRINKPWRISHSPQQRNPNS--------------human 47 VLCPSNSSQRVPLQAQK-LVSSHKPVQNQ---KQKQLQAT---SVPHPVSRPLNNTQKSKmus 46 VLCPSNS-QRVPSQAQK-LGAGQKPAP-------KQLPAA---SVPRPVSR-LNNPQKNExenla 49 ILGPSNVPQRVLAQAQKPILSSQKPTTQIPLRPATQGHQS---SKPQGPNENRNPQQTSHzebrafish 22 ----LNSRH-------------------------AELKKT---GNPE-----FQQTPFCWdrome 61 SAATTVRPATKPGLGGSNSIASSEGNNFQKPMVPSVKKTTSEFAAPAPVAPIKKPESLSKworm 10 --------------------------------------------------------VIDDyeast 47 -------------------------------------------KIPSPVREKLNRLPVNNhuman 100 QPLPSAPEN--NPEEE-LASKQK-NEESK--------KRQWALEDFEIGRPLGKGKFGNVmus 93 Q--PAASGN--DSEKE-QASLQK-TEDTK--------KRQWTLEDFDIGRPLGKGKFGNVxenla 106 SSTPNVEKK--GSTDQGKTSAVP-KEEGK--------KKQWCLEDFEIGRPLGKGKFGNVzebrafish 45 CGPLRVEMN--PDTHAVSGPGRV-PVKSN--------SKVLSIDDFDIGRPLGKGKFGNVdrome 121 QKPTAASSE--SSKELGAASSSAEKEKTKTETQPQKPKKTWELNNFDIGRLLGRGKFGNVworm 14 QKAEVISLT--EDSRPQRVDQAR--EESC-----------WSLDDFDVGRPLGKGKFGNVyeast 64 KKFLDMESSKIPSPIRKATSSKMIHENKK-----LPKFKSLSLDDFELGKKLGKGKFGKVhuman 148 YLAREKQSKFILALKVLFKAQLEKAGVEHQLRREVEIQSHLRHPNILRLYGYFHDATRVYmus 139 YLARERQSKFILALKVLFKTQLEKANVEHQLRREVEIQSHLRHPNILRLYGYFHDATRVYxenla 155 YLARERESKFILALKVLFKSQLEKAGVEHQLRREVEIQSHLRHPNILRLYGYFHDASRVYzebrafish 94 YLARERKLKVVIALKVLFKSQMVKEGVEHQLRREIEIQSHLRHPNILRFYNYFHDDTRVFdrome 179 YLAREKESQFVVALKVLFKRQIGESNVEHQVRREIEIQSHLRHPHILRLYAYFHDDVRIYworm 59 FISREKKTKRIIALKVLFKTQLLQLGVSHQLKREIEIQYHLRHPNILTLYGYFHDDKRVFyeast 119 YCVRHRSTGYICALKVMEKEEIIKYNLQKQFRREVEIQTSLNHPNLTKSYGYFHDEKRVY Amino acid sequence alignment of Aurora A kinase among species.

  6. Post-translational modification prediction • NetNGlyc - Prediction of N-glycosylation sites in human • proteins MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVLCPSNSSQRVPLQAQKLVSSHKPVQNQKQKQLQ 80 ATSVPHPVSRPLNNTQKSK QPLPSAPENNPEEELASKQKNEESKK ...................................................N............................ 80 ............................................. 160 SeqName Position Potential Jury agreement result Sequence 52 NSSQ 0.5574 (5/9) + Sequence 93 NNTQ 0.4741 (4/9) -

  7. No. Pos. Group Score 1 K119 ELASK QKNE ESKK  0.50 Post-translational modification prediction SignalP - Prediction of signal peptide cleavage sites: Not found HMMTOP - Prediction of transmembrane helices and topology of proteins (Hungarian Academy of Sciences): Not found • SUMOplot - Prediction of SUMO protein attachment sites HCA-Hydrophobic Cluster Analysis : Not found

  8. Post-translational modification prediction NetPhos - Prediction of Ser, Thr and Tyr phosphorylation sites in eukaryotic proteins Phosphorylation sites predicted: Ser: 4 Thr: 0 Tyr: 0 Serine predictions Name Pos Context Score Pred _________________________v_________________ Sequence 4 -MDRSKENC 0.045 . Sequence 10 ENCISGPVK 0.006 . Sequence 41 LPVNSGQAQ 0.125 . Sequence 51 VLCPSNSSQ 0.523 *S* Sequence 53 CPSNSSQRV 0.013 . Sequence 54 PSNSSQRVP 0.331 . Sequence 66 QKLVSSHKP 0.800 *S* Sequence 67 KLVSSHKPV 0.368 . Sequence 83 LQATSVPHP 0.056 . Sequence 89 PHPVSRPLN 0.007 . Sequence 98 NTQKSKQPL 0.323 . Sequence 104 QPLPSAPEN 0.634 *S* Sequence 116 EELASKQKN 0.352 . Sequence 123 KNEESKK-- 0.650 *S* Threonine predictions Name Pos Context Score Pred _________________________v_________________ Sequence 16 PVKATAPVG 0.036 . Sequence 28 RVLVTQQFP 0.011 . Sequence 82 QLQATSVPH 0.087 . Sequence 95 PLNNTQKSK 0.498 .

  9. Scansite: kinase site & binding domain profiles Src homology 3 group (SH3) Cortactin SH3 Gene Card HCLS1 Site Score Percentile Sequence SA P103 0.5524 0.278 % QKSKQPLPSAPENNP 0.946 Src SH3 Gene Card SRC Site Score Percentile Sequence SA P103 0.5116 0.823 % QKSKQPLPSAPENNP 0.946 Motifs scanned: AllStringency: Medium

  10. PDB code: 1MQ4 TITLE CRYSTAL STRUCTURE OF AURORA-A PROTEIN KINASE

  11. CONCLUSION • The Aurora Kinase A is very conserve throughout evolution. • Human Auroras A-C have a conserved kinase domain and • exhibit a relatively high sequence divergence at N terminal. • SUMO protein attachment sites. • Phosphorylation sites predicted (Ser: 4) • Possible binding domains: SH3 • Aurora-A is crystalized

  12. THANKS!!!

More Related