Mutational analysis of the phosphorylation sites of the Aie1 (aurora-C) kinase in vitro

被引:16
作者
Chen, SH [1 ]
Tang, TK [1 ]
机构
[1] Acad Sinica, Inst Biomed Sci, Taipei 115, Taiwan
关键词
D O I
10.1089/10445490252810302
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We previously reported two novel serine/threonine kinases, Aie1 (mouse) and AIE2 (human), both later referred to as aurora-C, a newly recognized member of the mitotic aurora kinase family. In the present study, we analyzed the phosphorylation sites of mouse Aiel by site-directed mutagenesis. Our results showed that protein kinase A (PKA) phosphoryIates Aiel at a threonine residue located at amino acid position 171. The T171A and T175A mutants, in which threonines located at residues 171 and 175 were replaced by alanines, revealed a significant increase in their kinase activities to phosphorylate ACS-1 (Aurora-C substrate 1). In contrast, the double mutant T171A-T175A showed impaired kinase activity. In addition, we had previously identified a PEST-like motif located at the N terminus of Aiel. Mutation analysis in the present study revealed that the quadruple mutant in which the PEST-like motif was mutated significantly abrogated Aiel kinase activity. This is the first report of the analysis of potential phosphorylation sites of mouse aurora-C in vitro.
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页码:41 / 46
页数:6
相关论文
共 21 条
[1]   PHOSPHORYLATION MODULATES CATALYTIC FUNCTION AND REGULATION IN THE CAMP-DEPENDENT PROTEIN-KINASE [J].
ADAMS, JA ;
MCGLONE, ML ;
GIBSON, R ;
TAYLOR, SS .
BIOCHEMISTRY, 1995, 34 (08) :2447-2454
[2]   Cloning of STK13, a third human protein kinase related to Drosophila aurora and budding yeast Ipl1 that maps on chromosome 19q13.3-ter [J].
Bernard, M ;
Sanseau, P ;
Henry, C ;
Couturier, A ;
Prigent, C .
GENOMICS, 1998, 53 (03) :406-409
[3]   A homologue of Drosophila aurora kinase is oncogenic and amplified in human colorectal cancers [J].
Bischoff, JR ;
Anderson, L ;
Zhu, YF ;
Mossie, K ;
Ng, L ;
Souza, B ;
Schryver, B ;
Flanagan, P ;
Clairvoyant, F ;
Ginther, C ;
Chan, CSM ;
Novotny, M ;
Slamon, DJ ;
Plowman, GD .
EMBO JOURNAL, 1998, 17 (11) :3052-3065
[4]   The Aurora/Ipi1p kinase family: regulators of chromosome segregation and cytokinesis [J].
Bischoff, JR ;
Plowman, GD .
TRENDS IN CELL BIOLOGY, 1999, 9 (11) :454-459
[5]  
Boyle WJ., 1991, METHOD ENZYMOL, V201, P110
[6]   REGULATION OF THE MICROTUBULE NUCLEATING ACTIVITY OF CENTROSOMES IN XENOPUS EGG EXTRACTS - ROLE OF CYCLIN-A-ASSOCIATED PROTEIN-KINASE [J].
BUENDIA, B ;
DRAETTA, G ;
KARSENTI, E .
JOURNAL OF CELL BIOLOGY, 1992, 116 (06) :1431-1442
[7]   TYPE-1 PROTEIN PHOSPHATASE ACTS IN OPPOSITION TO IPL1 PROTEIN-KINASE IN REGULATING YEAST CHROMOSOME SEGREGATION [J].
FRANCISCO, L ;
WANG, WF ;
CHAN, CSM .
MOLECULAR AND CELLULAR BIOLOGY, 1994, 14 (07) :4731-4740
[8]  
Giet R, 1999, J CELL SCI, V112, P3591
[9]   MUTATIONS IN AURORA PREVENT CENTROSOME SEPARATION LEADING TO THE FORMATION OF MONOPOLAR SPINDLES [J].
GLOVER, DM ;
LEIBOWITZ, MH ;
MCLEAN, DA ;
PARRY, H .
CELL, 1995, 81 (01) :95-105
[10]   Genomic organization, expression, and chromosome localization of a third aurora-related kinase gene, Aie1 [J].
Hu, HM ;
Chuang, CK ;
Lee, MJ ;
Tseng, TC ;
Tang, TK .
DNA AND CELL BIOLOGY, 2000, 19 (11) :679-688