Coupling of the RAS-MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase

Science

Volumn 273, Issue 5277, 1996, Pages 959-963

  Xing, Jun ^a   Ginty, David D ^b   Greenberg, Michael E ^a,c  

^a HARVARD MEDICAL SCHOOL   (United States)

^b JOHNS HOPKINS UNIVERSITY SCHOOL OF MEDICINE   (United States)

^c BOSTON CHILDREN S HOSPITAL   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CYCLIC AMP RESPONSIVE ELEMENT BINDING PROTEIN;

ARTICLE; ENZYME ANALYSIS; GENE ACTIVATION; GENE EXPRESSION REGULATION; GENETIC TRANSCRIPTION; IMMEDIATE EARLY GENE; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; REGULATORY MECHANISM;

EID: 0029789643     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5277.959     Document Type: Article

References (43)

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29. 222 with Glu residues.
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39. Abbreviations for the amino acid residues are as follows: A, Ala; 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; and Y, Tyr.
40. 2, and 1 mM DTT]. Washed immune complexes were used to phosphorylate CREBtide or recombinant CREB in kinase reactions as described (8).
41. Transfection of COS cells was done by the DEAE-dextran method [F. M. Ausubel et al., Eds., Current Protocols in Molecular Biology (Wiley, New York, 1994)]. Two days after transfection, cells were treated as described (Figs. 3 and 4). Cells were washed in ice-cold PBS and lysed in boiling SDS sample buffer for in-gel kinase assay or immunoblotting analysis. Alternatively, cells extracts were prepared and subjected to an immune complex kinase assay (26).
42. Assays for CAT and β-galactosidase were done as described [J. Sambrook et al., Eds., Molecular Cloning (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1989)].
43. We thank D. E. Moller and Y. Zhao for providing antibodies to RSK3, R. L. Erikson for murine RSK2 cDNA, E. G. Krebs for plasmids pCDNA3-MEK1(SE218/222) and pCDNA3-MEK1(KA97), N. K. Tonks and H. Sun for pSG5-MKP1, M. Z. Gilman for CMV-GAL4-CREB, S. H. Orkin for cloning vector pMT2, members of the Greenberg laboratory for critical reading of the manuscript, A. Bonni and Z. Xia for helpful discussions, and S. E. Kim and Y. Zhang for technical assistance. D.D.G. is a recipient of a Klingenstein Award in Neuroscience, an American Cancer Society Junior Faculty Research Award, and an Alfred Sloan Research Fellowship, Supported by National Institutes of Health grants NS34814-01 (D.D.G.) and CA43855 (M.E.G.), Mental Retardation Research Center grant NIH P30-HD18655, and an American Cancer Society Faculty Research Award (FRA-379) (M.E.G.).