Download Characterization of N-terminal of Aurora

The “deep” characterization
of Aurora A N-terminus
Isabel Peset Martin.
Vanessa Ferreira
Leire Mendizabal Oiarzabal
Human Aurora kinases A–C exhibit differential substrate affinity,
subcell localization and associated activities
Substrate
Cell localization
Effect of substrates-Aurora interaction
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; Rec8; 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
Paul D Andrews
Oncogene (2005)
DOMAIN STRUCTURE OF Aurora kinases A–C
AURORA A
AURORA B
AURORA C
Source
Domain
Start
End
PfamA
PKinase
133
383
SMART
STKc(hidden) 133
383
Source
Domain
Start
End
PfamB
Pfam-B 34136
1
62
SMART
STKc(hidden)
77
327
PfamA
PKinase
77
327
seg
Low Complexity (hidden) 204
215
Source
Domain
Start
End
PfamA
PKinase
43
293
SMART
STKc(hidden) 43
293
Amino acid sequence alignement of human
Aurora kinases A–C.
aurB
aurC
aurA
1 MAQKENSYPWPYGRQTAPSG-------------------LSTLPQRVLR----KEPVTPS
1 ----------------------------------------MSSPRAVVQ----LGKAQPA
1 MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVLCPSNSSQRVPLQ
aurB
aurC
aurA
38 ALVLMSRSN---------VQPTAAPG------------------------QKVMENSSGT
17 GEELATAN-----------QTAQQPS------------------------SPAM-----61 AQKLVSSHKPVQNQKQKQLQATSVPHPVSRPLNNTQKSKQPLPSAPENNPEEELASKQKN
aurB
aurC
aurA
65 PDILTRHFTIDDFEIGRPLGKGKFGNVYLAREKKSHFIVALKVLFKSQIEKEGVEHQLRR
36 -----RRLTVDDFEIGRPLGKGKFGNVYLARLKESHFIVALKVLFKSQIEKEGLEHQLRR
121 EESKKRQWALEDFEIGRPLGKGKFGNVYLAREKQSKFILALKVLFKAQLEKAGVEHQLRR
aurB
aurC
aurA
125 EIEIQAHLHHPNILRLYNYFYDRRRIYLILEYAPRGELYKELQKSCTFDEQRTATIMEEL
91 EIEIQAHLQHPNILRLYNYFHDARRVYLILEYAPRGELYKELQKSEKLDEQRTATIIEEL
181 EVEIQSHLRHPNILRLYGYFHDATRVYLILEYAPLGTVYRELQKLSKFDEQRTATYITEL
aurB
aurC
aurA
185 ADALMYCHGKKVIHRDIKPENLLLGLKGELKIADFGWSVHAPSLRRKTMCGTLDYLPPEM
151 ADALTYCHDKKVIHRDIKPENLLLGFRGEVKIADFGWSVHTPSLRRKTMCGTLDYLPPEM
241 ANALSYCHSKRVIHRDIKPENLLLGSAGELKIADFGWSVHAPSSRRTTLCGTLDYLPPEM
aurB
aurC
aurA
245 IEGRMHNEKVDLWCIGVLCYELLVGNPPFESASHNETYRRIVKVDLKFPASVPTGAQDLI
211 IEGRTYDEKVDLWCIGVLCYELLVGYPPFESASHSETYRRILKVDVRFPLSMPLGARDLI
301 IEGRMHDEKVDLWSLGVLCYEFLVGKPPFEANTYQETYKRISRVEFTFPDFVTEGARDLI
aurB
aurC
aurA
305 SKLLRHNPSERLPLAQVSAHPWVRANSRRVLPPSALQSVA--271 SRLLRYQPLERLPLAQILKHPWVQAHSRRVLPPCAQMAS---361 SRLLKHNPSQRPMLREVLEHPWITANSSKPSNCQNKESASKQS
Amino acid sequence alignment of
Aurora A kinase among species.
human
mus
xenla
zebrafish
drome
worm
yeast
1
1
1
1
1
1
1
------MDRS-KENCISGPVKATAPVGG-PKRVLVTQQFPCQNPLP------VNSGQAQR
human
------MDRC-KENCVSRPVKTTVPFG--PKRVLVTEQIPSQNLGS------ASSGQAQR
mus
------MERAVKENHKPSNVKIFHPMTEGAKRIPVNQPQSTQFRPP------GTAVSAQR
xenla
--------------------------------VFAVDREAEEKKHT------RTGGTLDzebrafish
MSHPSDHVLRPKENAPHRMPEKSAAVLNMQKNLLLGKKPNSENMAPDSKPLPGSSGALIR
drome
-------------------------------------MSGKENTAP-------------worm
MQRNSLVNIKLNANSPSKKTTTRPNTSRINKPWRISHSPQQRNPNS-------------yeast
208
199
215
154
239
119
179
LILEYAPLGTVYRELQK--LSKFDEQRTATYITELANALSYCHSKRVIHRDIKPENLLLG
LILEYAPLGTVYRELQK--LSKFDEQRTATYITELANALSYCHSKRVIHRDIKPENLLLG
LILDYAPGGELFRELQK--CTRFDDQRSAMYIKQLAEALLYCHSKKVIHRDIKPENLLLG
LILEYAPRGEMYKELQR--YGHFDDQRTATYMEEVSDALQYCHEKKVIHRDIKPENLLLG
LILEYAPQGTLFNALQAQPMKRFDERQSATYIQALCSALLYLHERDIIHRDIKPENLLLG
VILDYASRGELFNVLQSQPGHKVNEVIAGRFVRQLANALHYCHSKGVIHRDIKPENLLLD
LLMEYLVNGEMYKLLRL--HGPFNDILASDYIYQIANALDYMHKKNIIHRDIKPENILIG
human
mus
xenla
zebrafish
drome
worm
yeast
47
46
49
22
61
10
47
VLCPSNSSQRVPLQAQK-LVSSHKPVQNQ---KQKQLQAT---SVPHPVSRPLNNTQKSK
human
VLCPSNS-QRVPSQAQK-LGAGQKPAP-------KQLPAA---SVPRPVSR-LNNPQKNE
mus
ILGPSNVPQRVLAQAQKPILSSQKPTTQIPLRPATQGHQS---SKPQGPNENRNPQQTSH
xenla
----LNSRH-------------------------AELKKT---GNPE-----FQQTPFCW
zebrafish
SAATTVRPATKPGLGGSNSIASSEGNNFQKPMVPSVKKTTSEFAAPAPVAPIKKPESLSK
drome
--------------------------------------------------------VIDD
worm
-------------------------------------------KIPSPVREKLNRLPVNN
yeast
266
257
273
212
299
179
237
SAGELKIADFGWSVHAP-SSRRTTLCGTLDYLPPEMIEGRMHDEKVDLWSLGVLCYEFLV
SNGELKIADFGWSVHAP-SSRRTTMCGTLDYLPPEMIEGRMHDEKVDLWSLGVLCYEFLV
SNGELKIADFGWSVHAP-SSRRTTLCGTLDYLPPEMIEGRMHDETVDLWSLGVLCYEFLV
YRGELKIADFGWSVHAP-SLRRRTMCGTLDYLPPEMIEGHSHDEKVDLWSIGVLCYECLV
HKGVLKIADFGWSVHEP-NSMRMTLCGTVDYLPPEMVQGKPHTKNVDLWSLGVLCFELLV
SKLNLKLADFGWSVVAD-HSKRHTLCGTMDYLAPEMVSNQPHDFNVDIWAIGILLFEMLV
FNNVIKLTDFGWSIINPPENRRKTVCGTIDYLSPEMVESREYDHTIDAWALGVLAFELLT
human
mus
xenla
zebrafish
drome
worm
yeast
100
93
106
45
121
14
64
QPLPSAPEN--NPEEE-LASKQK-NEESK--------KRQWALEDFEIGRPLGKGKFGNV
human
Q--PAASGN--DSEKE-QASLQK-TEDTK--------KRQWTLEDFDIGRPLGKGKFGNV
mus
SSTPNVEKK--GSTDQGKTSAVP-KEEGK--------KKQWCLEDFEIGRPLGKGKFGNV
xenla
CGPLRVEMN--PDTHAVSGPGRV-PVKSN--------SKVLSIDDFDIGRPLGKGKFGNV
zebrafish
QKPTAASSE--SSKELGAASSSAEKEKTKTETQPQKPKKTWELNNFDIGRLLGRGKFGNV
drome
QKAEVISLT--EDSRPQRVDQAR--EESC-----------WSLDDFDVGRPLGKGKFGNV
worm
KKFLDMESSKIPSPIRKATSSKMIHENKK-----LPKFKSLSLDDFELGKKLGKGKFGKV
yeast
325
316
332
271
358
238
297
GKPPFEANTYQETYKRISRVEFTFPDFVTEGARDLISRLLKHNPSQRPMLREVLEHPWIT
GMPPFEAHTYQETYRRISRVEFTFPDFVTEGARDLISRLLKHNASQRLTLAEVLEHPWIK
GKPPFETDTHQETYRRISKVEFQYPPYVSEEARDLVSKLLKHNPNHRLPLKGVLEHPWII
GNPPFETRQHAETYKRITKVDLQFPKLVSEGARDLISKLLRHSPSMRLPLRSVMEHRGVK
GHAPFYSKNYDETYKKILKVDYKLPEHISKAASHLISKLLVLNPQHRLPLDQVMVHPWIL
GYAPFANQTGDKLIARIKECKIYIPSVVTDGAASLINAIIKKEPQERLPLVDIMAHPWIK
GAPPFEEEMKDTTYKRIAALDIKMPSNISQDAQDLILKLLKYDPKDRMRLGDVKMHPWIL
human
mus
xenla
zebrafish
drome
worm
yeast
148
139
155
94
179
59
119
YLAREKQSKFILALKVLFKAQLEKAGVEHQLRREVEIQSHLRHPNILRLYGYFHDATRVY
human
YLARERQSKFILALKVLFKTQLEKANVEHQLRREVEIQSHLRHPNILRLYGYFHDATRVY
mus
YLARERESKFILALKVLFKSQLEKAGVEHQLRREVEIQSHLRHPNILRLYGYFHDASRVY
xenla
YLARERKLKVVIALKVLFKSQMVKEGVEHQLRREIEIQSHLRHPNILRFYNYFHDDTRVF
zebrafish
YLAREKESQFVVALKVLFKRQIGESNVEHQVRREIEIQSHLRHPHILRLYAYFHDDVRIY
drome
FISREKKTKRIIALKVLFKTQLLQLGVSHQLKREIEIQYHLRHPNILTLYGYFHDDKRVF
worm
YCVRHRSTGYICALKVMEKEEIIKYNLQKQFRREVEIQTSLNHPNLTKSYGYFHDEKRVY
yeast
385
376
392
331
418
298
357
ANSSKP-SNCQNKESASKQS--------ANSSKPPTGHTSKEPTSKSS--------KNSQLK----KKDEPLPGAQ--------ANSRRVLPPVCSSDPH------------AHTQ------------------------EMKQREDIEVPLFISTLTKSSRNNSTANQ
RNKPFWENKRL------------------
Post-translational modification prediction
•NetNGlyc - Prediction of N-glycosylation sites in human
proteins
MDRSKENCISGPVKATAPVGGPKRVLVTQQFPCQNPLPVNSGQAQRVLCPSNSSQRVPLQAQKLVSSHKPVQNQKQKQLQ 80 ATSVPHPVSRPLNNTQKSK
QPLPSAPENNPEEELASKQKNEESKK ...................................................N............................ 80 ............................................. 160
SeqName
Sequence
Sequence
Position
52 NSSQ
93 NNTQ
Potential
0.5574
0.4741
Jury agreement
(5/9)
(4/9)
result
+
-
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
No.
1
Pos.
K119
Group
ELASK QKNE ESKK
HCA-Hydrophobic Cluster Analysis : Not found
Score
0.50
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
Threonine predictions
_________________________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*
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 .
•Scansite: kinase site & binding domain profiles
Motifs scanned: All
Stringency: Medium
Src homology 3 group (SH3)
Cortactin SH3
Gene Card HCLS1
Percentil
e
Site
Score
Sequence
SA
P103
0.5524 0.278 %
Src SH3
QKSKQPLPSAPENNP
Gene Card SRC
0.946
Site
Score
Percentil
e
Sequence
SA
P103
0.5116
0.823 %
QKSKQPLPSAPENNP
0.946
PDB code: 1MQ4
TITLE CRYSTAL STRUCTURE OF AURORA-A PROTEIN KINASE
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
THANKS!!!