Phosphorylation and activation of p70(s6k) by PDK1

Science

Volumn 279, Issue 5351, 1998, Pages 707-710

  Pullen, Nicholas ^a   Dennis, Patrick B ^a   Andjelkovic, Mirjana ^a   Dufner, Almut ^a   Kozma, Sara C ^a   Hemmings, Brian A ^a   Thomas, George ^a  

^a FRIEDRICH MIESCHER INSTITUTE FOR BIOMEDICAL RESEARCH   (Switzerland)

Author keywords

[No Author keywords available]

Indexed keywords

PHOSPHATIDYLINOSITOL KINASE; PROTEIN KINASE; 3 PHOSPHOINOSITIDE DEPENDENT PROTEIN KINASE; 3-PHOSPHOINOSITIDE-DEPENDENT PROTEIN KINASE; ANDROSTANE DERIVATIVE; CALMODULIN DEPENDENT PROTEIN KINASE IV; CALMODULIN-DEPENDENT PROTEIN KINASE IV; HYPERGLYCEMIC AGENT; INSULIN; ONCOPROTEIN; PHOSPHOTHREONINE; POLYENE; PROTEIN KINASE (CALCIUM,CALMODULIN); PROTEIN KINASE B; PROTEIN SERINE THREONINE KINASE; RAPAMYCIN; RECOMBINANT PROTEIN; S6 KINASE; WORTMANNIN;

ARTICLE; ENZYME ACTIVATION; ENZYME ACTIVE SITE; ENZYME PHOSPHORYLATION; PRIORITY JOURNAL; AMINO ACID SEQUENCE; ANIMAL; BINDING SITE; CATALYSIS; CELL LINE; METABOLISM; MOLECULAR GENETICS; PHOSPHORYLATION;

AMINO ACID SEQUENCE; ANDROSTADIENES; ANIMALS; BINDING SITES; CA(2+)-CALMODULIN DEPENDENT PROTEIN KINASE; CATALYSIS; CELL LINE; ENZYME ACTIVATION; INSULIN; INSULIN ANTAGONISTS; MOLECULAR SEQUENCE DATA; PHOSPHORYLATION; PHOSPHOTHREONINE; POLYENES; PROTEIN-SERINE-THREONINE KINASES; PROTO-ONCOGENE PROTEINS; PROTO-ONCOGENE PROTEINS C-AKT; RECOMBINANT PROTEINS; RIBOSOMAL PROTEIN S6 KINASES; SIROLIMUS;

EID: 0032578998     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.279.5351.707     Document Type: Article

References (23)

1. H. B. J. Jeffries et al., EMBO J. 16, 3693 (1997).
2. O. Meyuhas, D. Avni, S. Shama, in Translational Control, J. W. B. Hershey, M. B. Mathews, N. Sonenberg, Eds. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1996).
3. J. Chung, T. C. Grammer, K. P. Lemon, A. Kazlauskas, J. Blenis, Nature 370, 71 (1994); X. F. Ming et al., ibid. 371, 426 (1994).
4. J. Chung, T. C. Grammer, K. P. Lemon, A. Kazlauskas, J. Blenis, Nature 370, 71 (1994); X. F. Ming et al., ibid. 371, 426 (1994).
5. N. Pullen and G. Thomas, FEBS Lett. 410, 78 (1997).
6. B. A. Moser et al., Mol. Cell. Biol. 17, 5648 (1997).
7. S. K. Hanks and T. Hunter, in The Protein Kinase Facts Book: Protein-Serine Kinases, G. Hardie and S. Hanks, Eds. (Academic Press, London, 1995).
8. D. R. Alessi et al., Curr. Biol. 7, 261 (1997); D. Stokoe et al., Science 277, 567 (1997).
9. D. R. Alessi et al., Curr. Biol. 7, 261 (1997); D. Stokoe et al., Science 277, 567 (1997).
10. D. R. Alessi et al., Curr. Biol. 7, 776 (1997).
11. P. B. Dennis, N. Pullen, S. C. Kozma, G. Thomas, Mol. Cell. Biol. 16, 6242 (1996).
12. The cDNA encoding PDK1 (amino acids 52 through 556) was isolated with use of the sequence derived from the Drosophila homolog, DSTPK61/Y07908. Using the Drosophila sequence, we screened human expressed sequence tag (EST) databases and assembled an open reading frame covering amino acids 52 through 556. This open reading frame was confirmed by screening the Institute for Genomic Research (TIGR) EST database. The cDNA was obtained by polymerase chain reaction (PCR) from a placenta library (Clonetech) with use of the following oligonucleotides: 5′-CGC ATG GAC GGC ACT GCA GCC GAG CCT CG and 3'′-CGC AGG CCA CGT CAC TGC ACA GCG GCG. After confirmation of the sequence, a Myc epitope (EQKLISEEDL) was added by a second round of PCR (5'′-CCC GGT ACC GCC ATG GAA CAG AAA CTC ATC TCT GAA GAG GAT CTG GAC GGC ACT GCA GCC GAG CCT CG and 3′-CCC TCT AGA TCA CTG CAC AGC GGC GTC CGG GTG GC). The cDNA was subdoned as a Kpn I-Xba I fragment into pCMV5 and used in subsequent experiments. We generated Myc-tagged PDK1-KD (PDK1Q61) using Quickchange (Stratagene) with 5′-CGC TTC TCC AGA ATC TGA ATC GCA TAT TCT CTG G and 3′-CCA GAG AAT ATG CGA TTC AGA TTC TGG AGA AGC G oligonucleotides on the pCMV5-PDK1 construct.
13. M. Andjelkovich et al., Proc. Natl. Acad. Sci. U.S.A. 93, 5699 (1996).
14. mapk activities were assayed as described in (1, 19).
15. P. B. Dennis, N. Pullen, S. C. Kozma, G. Thomas, unpublished observations.
16. R. B. Pearson et al., EMBO J, 14, 5279 (1995).
17. 3E.
18. H. Tokumitsu, H. Enslen, T. R. Soderling, J. Biol. Chem. 270, 19320 (1995).
19. Q.-P. Weng et al., Proc. Natl. Acad. Sci. U.S.A. 92, 5744 (1995).
20. Further PDK1-related cDNAs have been isolated with use of the PDK1 cDNA isolated from the placenta library (B. A. Hemmings, unpublished observations).
21. D. A. Cross, D. R. Alessi, P. Cohen, M. Andjelkovic, B. A. Hemmings, Nature 378, 785 (1995).
22. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
23. This work is supported by a Human Frontier Science Program Organisation (HFSPO) Fellowship to N.P. and P.B.D. and grants from the European Economic Community and HFSPO to G.T. We acknowledge P. Cron for the initial isolation and characterization of the PDK1 cDNA, D. Brickey and T. Soderling (Vollum Institute, Portland, OR) for purified CaMK IV and CaMK kinase, M. Rothnie for preparing figures, and M. Frech for help and advice.