The protein kinase p90 Rsk as an essential mediator of cytostatic factor activity

Science

Volumn 286, Issue 5443, 1999, Pages 1362-1365

  Bhatt, Ramesh R ^a   Ferrell Jr , James E ^a  

^a STANFORD UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CYTOSTATIC AGENT; MITOGEN ACTIVATED PROTEIN KINASE; PROTEIN KINASE;

ARTICLE; CELL CYCLE; ENZYME ACTIVATION; ENZYME ANALYSIS; ENZYME INACTIVATION; ENZYME PHOSPHORYLATION; MITOSIS INHIBITION; NONHUMAN; PRIORITY JOURNAL; XENOPUS LAEVIS;

ANIMALS; CDC2 PROTEIN KINASE; CELL EXTRACTS; ENZYME ACTIVATION; MAP KINASE KINASE 1; MAP KINASE SIGNALING SYSTEM; MITOGEN-ACTIVATED PROTEIN KINASE 1; MITOGEN-ACTIVATED PROTEIN KINASE KINASES; MITOSIS; MOLECULAR SEQUENCE DATA; OVUM; PROTEIN-SERINE-THREONINE KINASES; PROTO-ONCOGENE PROTEINS C-MOS; RIBOSOMAL PROTEIN S6 KINASES; XENOPUS LAEVIS;

EID: 0032695982     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5443.1362     Document Type: Article

References (40)

1. Y. Masui and C. L. Markert, J. Exp. Zool. 177, 129 (1971).
2. J. L. Maller, Trends Biochem. Sci. 20, 524 (1995).
3. N. Sagata, Bioessays 19, 13 (1997).
4. _, N. Sagata, N. Watanabe, G. F. Vande Woude, Y. Ikawa, Nature 342, 512 (1989).
5. N. Yew, M. L. Mellini, G. F. Vande Woude, Nature 355, 649 (1992).
6. O. Haccard et al., Science 262, 1262 (1993).
7. A. Abrieu et al., J. Cell Sci. 109, 239 (1996).
8. A. Abrieu, D. Fisher, M. N. Simon, M. Doree, A. Picard, EMBO J. 16, 6407 (1997); S. A. Walter, T. M. Guadagno, J. E. Ferrell Jr., Mol. Biol. Cell 8, 2157 (1997); M. S. Murakami and G. F. Vande Woude, Development 125, 237 (1998); J. C. Bitangcol et al., Mol. Biol. Cell 9, 451 (1998).
9. A. Abrieu, D. Fisher, M. N. Simon, M. Doree, A. Picard, EMBO J. 16, 6407 (1997); S. A. Walter, T. M. Guadagno, J. E. Ferrell Jr., Mol. Biol. Cell 8, 2157 (1997); M. S. Murakami and G. F. Vande Woude, Development 125, 237 (1998); J. C. Bitangcol et al., Mol. Biol. Cell 9, 451 (1998).
10. A. Abrieu, D. Fisher, M. N. Simon, M. Doree, A. Picard, EMBO J. 16, 6407 (1997); S. A. Walter, T. M. Guadagno, J. E. Ferrell Jr., Mol. Biol. Cell 8, 2157 (1997); M. S. Murakami and G. F. Vande Woude, Development 125, 237 (1998); J. C. Bitangcol et al., Mol. Biol. Cell 9, 451 (1998).
11. A. Abrieu, D. Fisher, M. N. Simon, M. Doree, A. Picard, EMBO J. 16, 6407 (1997); S. A. Walter, T. M. Guadagno, J. E. Ferrell Jr., Mol. Biol. Cell 8, 2157 (1997); M. S. Murakami and G. F. Vande Woude, Development 125, 237 (1998); J. C. Bitangcol et al., Mol. Biol. Cell 9, 451 (1998).
12. H. Kosako, Y. Gotoh, E. Nishida, J. Biol. Chem. 269, 28354 (1994).
13. D. A. Cross and C. Smythe, Exp. Cell Res. 241, 12 (1998).
14. W. H. Colledge, M. B. Carlton, G. B. Udy, M. J. Evans, Nature 370, 65 (1994); N. Hashimoto et al., Nature 370, 68 (1994).
15. W. H. Colledge, M. B. Carlton, G. B. Udy, M. J. Evans, Nature 370, 65 (1994); N. Hashimoto et al., Nature 370, 68 (1994).
16. T. M. Guadagno and J. E. Ferrell Jr., Science 282, 1312 (1998).
17. A. S. Chau and E. K. Shibuya, Biol. Cell 90, 565 (1998).
18. J. Blenis, Proc. Natl. Acad. Sci. U.S.A. 90, 5889 (1993).
19. V. M. Rivera et al., Mol. Cell. Biol. 13, 6260 (1993); R. H. Chen, C. Abate, J. Blenis, Proc. Natl. Acad. Sci. U.S.A. 90, 10952 (1993); J. Xing, D. D. Ginty, M. E. Greenberg, Science 273, 959 (1996); D. De Cesare, S. Jacquot, A. Hanauer, P. Sassone-Corsi, Proc. Natl. Acad. Sci. U.S.A. 95, 12202 (1998).
20. V. M. Rivera et al., Mol. Cell. Biol. 13, 6260 (1993); R. H. Chen, C. Abate, J. Blenis, Proc. Natl. Acad. Sci. U.S.A. 90, 10952 (1993); J. Xing, D. D. Ginty, M. E. Greenberg, Science 273, 959 (1996); D. De Cesare, S. Jacquot, A. Hanauer, P. Sassone-Corsi, Proc. Natl. Acad. Sci. U.S.A. 95, 12202 (1998).
21. V. M. Rivera et al., Mol. Cell. Biol. 13, 6260 (1993); R. H. Chen, C. Abate, J. Blenis, Proc. Natl. Acad. Sci. U.S.A. 90, 10952 (1993); J. Xing, D. D. Ginty, M. E. Greenberg, Science 273, 959 (1996); D. De Cesare, S. Jacquot, A. Hanauer, P. Sassone-Corsi, Proc. Natl. Acad. Sci. U.S.A. 95, 12202 (1998).
22. V. M. Rivera et al., Mol. Cell. Biol. 13, 6260 (1993); R. H. Chen, C. Abate, J. Blenis, Proc. Natl. Acad. Sci. U.S.A. 90, 10952 (1993); J. Xing, D. D. Ginty, M. E. Greenberg, Science 273, 959 (1996); D. De Cesare, S. Jacquot, A. Hanauer, P. Sassone-Corsi, Proc. Natl. Acad. Sci. U.S.A. 95, 12202 (1998).
23. E. Trivier et al., Nature 384, 567 (1996); F. Abidi et al., Eur. J. Hum. Genet. 7, 20 (1999).
24. E. Trivier et al., Nature 384, 567 (1996); F. Abidi et al., Eur. J. Hum. Genet. 7, 20 (1999).
25. E. Erikson and J. L. Maller, Proc. Natl. Acad. Sci. U.S.A. 82, 742 (1985); _, R. Erikson, Methods Enzymol. 200, 252 (1991); K.-M. Hsiao. S.-y. Chou, S.-J. Shih, J. E. Ferrell Jr., Proc. Natl. Acad. Sci. U.S.A. 91, 5480 (1994); A. Palmer, A. C. Gavin, A. R. Nebreda, EMBO J. 17, 5037 (1998).
26. E. Erikson and J. L. Maller, Proc. Natl. Acad. Sci. U.S.A. 82, 742 (1985); _, R. Erikson, Methods Enzymol. 200, 252 (1991); K.-M. Hsiao. S.-y. Chou, S.-J. Shih, J. E. Ferrell Jr., Proc. Natl. Acad. Sci. U.S.A. 91, 5480 (1994); A. Palmer, A. C. Gavin, A. R. Nebreda, EMBO J. 17, 5037 (1998).
27. E. Erikson and J. L. Maller, Proc. Natl. Acad. Sci. U.S.A. 82, 742 (1985); _, R. Erikson, Methods Enzymol. 200, 252 (1991); K.-M. Hsiao. S.-y. Chou, S.-J. Shih, J. E. Ferrell Jr., Proc. Natl. Acad. Sci. U.S.A. 91, 5480 (1994); A. Palmer, A. C. Gavin, A. R. Nebreda, EMBO J. 17, 5037 (1998).
28. E. Erikson and J. L. Maller, Proc. Natl. Acad. Sci. U.S.A. 82, 742 (1985); _, R. Erikson, Methods Enzymol. 200, 252 (1991); K.-M. Hsiao. S.-y. Chou, S.-J. Shih, J. E. Ferrell Jr., Proc. Natl. Acad. Sci. U.S.A. 91, 5480 (1994); A. Palmer, A. C. Gavin, A. R. Nebreda, EMBO J. 17, 5037 (1998).
29. Xenopus Rsk-2 is 92% identical to human Rsk-2 (ISPK-1) and 79% identical to Xenopus Rsk-1. From quantitative immunoblotting experiments with purified recombinant Rsk-1 and Rsk-2 proteins as standards, we have estimated the oocyte concentration of Rsk-1 to be 2 to 6 nM and Rsk-2 to be ∼100 nM. The Xenopus Rsk-2 sequence has been submitted to the National Center for Biotechnology Information (accession number AF165162).
30. 2 arrest in response to added Mos (7, 8), making the extracts better suited for examining Mos-induced mitotic arrests. CSF extracts were prepared as described (28).
31. J. Minshull, H. Sun, N. K. Tonks, A. W. Murray, Cell 79, 475 (1994).
32. Mos was expressed in bacteria as a maltose-binding protein (malE) fusion (5). The malE-Mos fusion protein was purified as described (5).
33. R. R. Bhatt and J. E. Ferrell Jr., unpublished data.
34. Rsk-2 immunodepletion was accomplished by incubating extracts for 75 min on ice with antibodies to Rsk-2 (Santa Cruz Biotechnology) that had been prebound to protein G agarose beads (Gibco-BRL). Mock depletions were carried out similarly, with the antibody omitted.
35. Immunodepletion of Rsk-1 yielded extracts that still underwent a CSF arrest in response to Mos. Thus, Rsk-1 appears not to be necessary for CSF arrest.
36. 6-tagged Rsk-2 proteins (with His tag inserted directly after the putative translation initiation methionine codon) were produced in Sf9 cells and purified to homogeneity by nickel chelate chromatography. Catalytically inactive Rsk-2 (KR Rsk-2) was engineered by substituting arginine for lysine at residue 97.
37. We also titrated different concentrations of Rsk-1 or Rsk-2 into Rsk-2-depleted extracts. We found that 100 nM Rsk-1 or Rsk-2 was sufficient to restore the arrest, but 20 nM, 4 nM, and 1 nM were insufficient. These findings indicate that the Rsks are at least partially interchangeable, but by virtue of its higher normal concentration (18) Rsk-2 is the more important Rsk in this this context Our results fit well with the recent finding that a gain-of-function form of Rsk-1 can cause CSF arrest [S. G. Gross, M. S. Schwab, A. L. Lewellyn, J. L. Maller, Science 286, 1365 (1999)].
38. M. Zecevic et al., J. Cell Biol. 142, 1547 (1998).
39. A. W. Murray, Methods Cell Biol. 36, 581 (1991).
40. We thank M. Murakami and G. Vande Woude for Mos plasmkis, C.-Y. F. Huang for preparing Mos protein, and M. L. Sohaskey and S. A. Walter for comments on the manuscript Supported by a grant from NIH (CM46383) and a National Research Service Award Predoctoral Fellowship (GM16415).