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Volumn 278, Issue 5339, 1997, Pages 860-866

IKK-1 and IKK-2: Cytokine-activated IκB kinases essential for NF-κB activation

Author keywords

[No Author keywords available]

Indexed keywords

HELIX LOOP HELIX PROTEIN; IMMUNOGLOBULIN ENHANCER BINDING PROTEIN; LEUCINE ZIPPER PROTEIN;

EID: 0030685825     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5339.860     Document Type: Article
Times cited : (1872)

References (53)
  • 1
    • 0028971289 scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1995) Genes Dev. , vol.9 , pp. 2723
    • Verma, I.M.1    Stevenson, J.K.2    Schwarz, E.M.3    Van Antwerp, D.4    Miyamoto, S.5
  • 2
    • 0028174061 scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1994) Annu. Rev. Immunol. , vol.12 , pp. 141
    • Baeuerle, P.1    Henkel, T.2
  • 3
    • 0000640646 scopus 로고    scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1996) Drug Discovery Today , vol.1 , pp. 151
    • Manning, A.1
  • 4
    • 0030615201 scopus 로고    scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1997) N. Engl. J. Med. , vol.336 , pp. 1066
    • Barnes, P.J.1    Karin, M.2
  • 5
    • 0030271387 scopus 로고    scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1996) Cell , vol.87 , pp. 13
    • Baeuerle, P.A.1    Baltimore, D.2
  • 6
    • 0030898311 scopus 로고    scopus 로고
    • I. M. Verma, J. K. Stevenson, E. M. Schwarz, D. Van Antwerp, S. Miyamoto, Genes Dev. 9, 2723 (1995); P. Baeuerle and T. Henkel, Annu. Rev. Immunol. 12, 141 (1994); A. Manning, Drug Discovery Today 1, 151 (1996); P. J. Barnes and M. Karin, N. Engl. J. Med. 336, 1066 (1997); P. A. Baeuerle and D. Baltimore, Cell 87, 13 (1996); V. R. Baichwal and P. A. Baeuerle, Curr. Biol. 7, 94 (1997).
    • (1997) Curr. Biol. , vol.7 , pp. 94
    • Baichwal, V.R.1    Baeuerle, P.A.2
  • 8
    • 0024759789 scopus 로고
    • P. A. Baeuerle and D. Baltimore, Cell 53, 211 (1988); _, Genes Dev. 3, 1689 (1989).
    • (1989) Genes Dev. , vol.3 , pp. 1689
  • 10
    • 0029865142 scopus 로고    scopus 로고
    • J. E. Thompson, R. J. Philips, H. Erdjument-Bromage, P. Tempst, S. Ghosh, Cell 80, 573 (1995); Z. Cao, W. Henzel, X. Gao, Science 271, 1128 (1996); U. Siebenlist, G. Franzoso, K. Brown, Annu. Rev. Cell Biol. 10, 405 (1994).
    • (1996) Science , vol.271 , pp. 1128
    • Cao, Z.1    Henzel, W.2    Gao, X.3
  • 11
    • 0028170816 scopus 로고
    • J. E. Thompson, R. J. Philips, H. Erdjument-Bromage, P. Tempst, S. Ghosh, Cell 80, 573 (1995); Z. Cao, W. Henzel, X. Gao, Science 271, 1128 (1996); U. Siebenlist, G. Franzoso, K. Brown, Annu. Rev. Cell Biol. 10, 405 (1994).
    • (1994) Annu. Rev. Cell Biol. , vol.10 , pp. 405
    • Siebenlist, U.1    Franzoso, G.2    Brown, K.3
  • 12
    • 0027332498 scopus 로고
    • A. A. Beg and A. S. Baldwin Jr., Genes Dev. 7, 2064 (1993); T. D. Gilmore and P. J. Morin, Trends Genet. 9, 427 (1993); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); S. Haskill et al., Cell 65, 1281 (1991).
    • (1993) Genes Dev. , vol.7 , pp. 2064
    • Beg, A.A.1    Baldwin Jr., A.S.2
  • 13
    • 0027333044 scopus 로고
    • A. A. Beg and A. S. Baldwin Jr., Genes Dev. 7, 2064 (1993); T. D. Gilmore and P. J. Morin, Trends Genet. 9, 427 (1993); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); S. Haskill et al., Cell 65, 1281 (1991).
    • (1993) Trends Genet. , vol.9 , pp. 427
    • Gilmore, T.D.1    Morin, P.J.2
  • 14
    • 0027295420 scopus 로고
    • A. A. Beg and A. S. Baldwin Jr., Genes Dev. 7, 2064 (1993); T. D. Gilmore and P. J. Morin, Trends Genet. 9, 427 (1993); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); S. Haskill et al., Cell 65, 1281 (1991).
    • (1993) Mol. Cell. Biol. , vol.13 , pp. 4770
    • Diaz-Meco, M.T.1
  • 15
    • 0025812292 scopus 로고
    • A. A. Beg and A. S. Baldwin Jr., Genes Dev. 7, 2064 (1993); T. D. Gilmore and P. J. Morin, Trends Genet. 9, 427 (1993); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); S. Haskill et al., Cell 65, 1281 (1991).
    • (1991) Cell , vol.65 , pp. 1281
    • Haskill, S.1
  • 16
    • 0026783210 scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1992) Genes Dev. , vol.6 , pp. 1899
    • Beg, A.A.1
  • 17
    • 0028985191 scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1995) Mol. Cell. Biol. , vol.15 , pp. 1294
    • Alkalay, I.1
  • 18
    • 0028985190 scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1995) Mol. Cell. Biol. , pp. 1302
    • DiDonato, J.A.1    Mercurio, F.2    Karin, M.3
  • 19
    • 0026663539 scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1992) Annu. Rev. Biochem. , vol.61 , pp. 761
    • Hershko, A.1    Ciechanover, A.2
  • 20
    • 0028972488 scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1995) Proc. Natl. Acad. Sci. U.S.A. , vol.92 , pp. 11259
    • Scherer, D.C.1    Brockman, J.2    Chen, Z.3    Maniatis, T.4    Ballard, D.5
  • 21
    • 0029665080 scopus 로고    scopus 로고
    • A. A. Beg et al., Genes Dev. 6, 1899 (1992); I. Alkalay et al., Mol. Cell. Biol. 15, 1294 (1995); J. A. DiDonato, F. Mercurio, M. Karin, ibid., p. 1302 (1995); A. Hershko and A. Ciechanover, Annu. Rev. Biochem. 61, 761 (1992); D. C. Scherer, J. Brockman, Z. Chen, T. Maniatis, D. Ballard, Proc. Natl. Acad. Sci. U.S.A. 92, 11259 (1995); M. Roff et al., J. Biol. Chem. 271, 7844 (1996).
    • (1996) J. Biol. Chem. , vol.271 , pp. 7844
    • Roff, M.1
  • 22
    • 0028986075 scopus 로고
    • K. Brown, S. Gerstberger, L. Carlson, G. Franzoso, U. Siebenlist, Science 267, 1485 (1995); E. B. M. Traenckner et al., EMBO J. 14, 2876 (1995); S. T. Whiteside et al., Mol. Cell. Biol. 15, 5339 (1995); J. A. DiDonato et al., ibid. 16, 1295 (1996).
    • (1995) Science , vol.267 , pp. 1485
    • Brown, K.1    Gerstberger, S.2    Carlson, L.3    Franzoso, G.4    Siebenlist, U.5
  • 23
    • 0028978032 scopus 로고
    • K. Brown, S. Gerstberger, L. Carlson, G. Franzoso, U. Siebenlist, Science 267, 1485 (1995); E. B. M. Traenckner et al., EMBO J. 14, 2876 (1995); S. T. Whiteside et al., Mol. Cell. Biol. 15, 5339 (1995); J. A. DiDonato et al., ibid. 16, 1295 (1996).
    • (1995) EMBO J. , vol.14 , pp. 2876
    • Traenckner, E.B.M.1
  • 24
    • 0029122799 scopus 로고
    • K. Brown, S. Gerstberger, L. Carlson, G. Franzoso, U. Siebenlist, Science 267, 1485 (1995); E. B. M. Traenckner et al., EMBO J. 14, 2876 (1995); S. T. Whiteside et al., Mol. Cell. Biol. 15, 5339 (1995); J. A. DiDonato et al., ibid. 16, 1295 (1996).
    • (1995) Mol. Cell. Biol. , vol.15 , pp. 5339
    • Whiteside, S.T.1
  • 25
    • 0029670083 scopus 로고    scopus 로고
    • K. Brown, S. Gerstberger, L. Carlson, G. Franzoso, U. Siebenlist, Science 267, 1485 (1995); E. B. M. Traenckner et al., EMBO J. 14, 2876 (1995); S. T. Whiteside et al., Mol. Cell. Biol. 15, 5339 (1995); J. A. DiDonato et al., ibid. 16, 1295 (1996).
    • (1996) Mol. Cell. Biol. , vol.16 , pp. 1295
    • DiDonato, J.A.1
  • 26
    • 1842295012 scopus 로고    scopus 로고
    • note
    • 4, 1 mM benzamidine, 2 μM PMSF, aprotinin at 10 μg/ml, leupeptin at 1 μg/ml, pepstatin at 1 μg/ml, 1 mM DTT], and the kinase was eluted in a gradient to PS buffer with no ammonium sulfate. Fractions containing IκB kinase activity were subjected to immunoblot analysis with anti-RelA, anti-IκB-β, and anti-MKP-1 (Santa Cruz Biotechnology, Inc.)or anti-MEKK-1 or anti-P-Tyr (Upstate Biotechnology).
  • 27
    • 1842416966 scopus 로고    scopus 로고
    • note
    • 4, 1 mM benzamidine, 2 μM PMSF, aprotinin at 10 μg/ml, leupeptin at 1 μg/ml, pepstatin at 1 μg/ml, 1 mM DTT] at 30°C for 30 to 60 min in the presence of the indicated substrate. The kinase reaction was stopped by addition of 6× SDS-polyacrylamide gel electrophoresis (PAGE) sample buffer, subjected to SDS-PAGE analysis, and visualized by autoradiography. GST-IκB substrates were expressed and purified from Escherichia coli with glutathione Sepharose 4B beads. His-tagged RelA-streptavidin-tagged IκB-α WT and His-tagged Rel A-streptavidin-tagged IκB-α (S32A, S36A) mutant complexes were purified from Sf9 cells coinfected with the indicated baculoviral expression constructs.
  • 30
    • 0031285250 scopus 로고    scopus 로고
    • F. S. Lee, J. Hagler, Z. J. Chen, T. Maniatis, ibid. 88, 213 (1997); M. Hirano et al., J. Biol. Chem. 271, 13234 (1996); N. L. Malinin, M. P. Bolin, A. V. Kovalenko, D. Wallach, Nature 385, 540 (1997).
    • (1997) Cell , vol.88 , pp. 213
    • Lee, F.S.1    Hagler, J.2    Chen, Z.J.3    Maniatis, T.4
  • 31
    • 0029947996 scopus 로고    scopus 로고
    • F. S. Lee, J. Hagler, Z. J. Chen, T. Maniatis, ibid. 88, 213 (1997); M. Hirano et al., J. Biol. Chem. 271, 13234 (1996); N. L. Malinin, M. P. Bolin, A. V. Kovalenko, D. Wallach, Nature 385, 540 (1997).
    • (1996) J. Biol. Chem. , vol.271 , pp. 13234
    • Hirano, M.1
  • 32
    • 0031017618 scopus 로고    scopus 로고
    • F. S. Lee, J. Hagler, Z. J. Chen, T. Maniatis, ibid. 88, 213 (1997); M. Hirano et al., J. Biol. Chem. 271, 13234 (1996); N. L. Malinin, M. P. Bolin, A. V. Kovalenko, D. Wallach, Nature 385, 540 (1997).
    • (1997) Nature , vol.385 , pp. 540
    • Malinin, N.L.1    Bolin, M.P.2    Kovalenko, A.V.3    Wallach, D.4
  • 33
    • 1842299149 scopus 로고    scopus 로고
    • note
    • 4, 1 mM benzamidine, 2 μM PMSF, aprotinin at 10 μg/ml, leupeptin at 1 μg/ml, pepstatin at 1 μg/ml, 1 mM DTT] or up to 3.5 M urea.
  • 34
    • 1842412921 scopus 로고    scopus 로고
    • note
    • 32P was detected by scintillation counting. Less than 10% of the ATP was consumed in the reaction.
  • 35
    • 1842334327 scopus 로고    scopus 로고
    • note
    • For large-scale IKK signalsome purification, HeLa S3 cells were stimulated for 7 min with TNF-α at 20 ng/ml (R&D Systems) and harvested; whole-cell lysate was prepared (1.2 g of total protein) and about 5 mg of anti-MKP-1 (Santa Cruz Biotechnology, Inc.) was added and incubated at 4°C for 2 hours with gentle rotation. Then 50 ml of protein A-agarose (Calbiochem) was added and incubated for 2 hours. The immunoprecipitate was then sequentially washed four times with PD buffer, two times with RIPA buffer, two times with PD buffer, once with 3.5 M urea-PD buffer, and three times with PD buffer. The immunoprecipitate was then made into a thick slurry by adding 10 ml of PD buffer and 25 mg of the MKP-1 peptide to which the antibody was generated (Santa Cruz Biotechnology, Inc.); then it was incubated overnight at 4°C with gentle rotation. Salt was removed from the eluted protein on PD10 columns (Pharmacia) equilibrated with 50 mM Q buffer, and eluate was chromatographed on a Mono Q column (Pharmacia). Fractions containing IκB kinase activity were pooled, concentrated, and subjected to preparative SDS-PAGE; protein bands were visualized with colloidal blue stain (Novex) and the bands were excised and sequenced.
  • 36
    • 1842295546 scopus 로고    scopus 로고
    • note
    • Coomassie blue-stained bands were excised and digested with trypsin as described. A small portion of the supernatant was removed for analysis by MALDI peptide mapping as described (16). The program PeptideSearch (EMBL, Heidelberg) was used to compare the peptide mass map from the IKK-1 band with a protein sequence database. Eight measured peptide masses matched those calculated for peptides from CHUK within 30 ppm (18). The peptide mass map of the IKK-2 band did not result in a clear identification and therefore the sample was subject-ed to nanoelectrospray mass spectrometry (25). The peptide mixture was micropurified on a capillary containing 50 nl of Poros R2 resin (Perseptive Biosystems, Framingham, MA). The peptides were washed and then step-eluted with 0.5 μl of 50% MeOH in 5% formic acid into a nanoelectrospray needle. This needle was transferred to an APIII mass spectrometer (Perkin-Elmer, Sciex, Toronto, Canada) and the sample was sprayed for about 20 min. During this time, peptide ions apparent from the mass spectrum were selected and isolated and then fragmented in the collision chamber of the mass spectrometer. From the tandem mass spectra, short stretches of sequence were assembled into peptide sequence tags (18) and compared with a protein sequence database or an EST database by using PeptideSearch. Three peptides matched the IKK-1 sequence. A1, IIDLGYAK; A2, VEVALSNIK; A3, SIQLDLER. Three other peptides matched human EST sequences in the EST database: B1, ALELLPK; B2, VIYTQLSK; B6, LLLQAIQSFEK. These three sequences all match EST clone AA326115. The peptide B4 with the sequence LGTGGFGNVIR was found in clone R06591. After the full-length IKK-2 sequence was obtained (19), two more peptides (B3, ALDDILNLK; B5, DLK-PENIVLQQGEQR) were found in the sequence. Peptide A1 is present in both IKK-1 and IKK-2 sequences.
  • 39
    • 0028575316 scopus 로고
    • M. Mann and M. S. Wilm, Anal. Chem. 66, 4390 (1994); M. Wilm et al., Nature 379, 466 (1996); M. Mann, Trends Biol. Sci. 21, 494 (1996).
    • (1994) Anal. Chem. , vol.66 , pp. 4390
    • Mann, M.1    Wilm, M.S.2
  • 40
    • 0030026070 scopus 로고    scopus 로고
    • M. Mann and M. S. Wilm, Anal. Chem. 66, 4390 (1994); M. Wilm et al., Nature 379, 466 (1996); M. Mann, Trends Biol. Sci. 21, 494 (1996).
    • (1996) Nature , vol.379 , pp. 466
    • Wilm, M.1
  • 41
    • 0030448246 scopus 로고    scopus 로고
    • M. Mann and M. S. Wilm, Anal. Chem. 66, 4390 (1994); M. Wilm et al., Nature 379, 466 (1996); M. Mann, Trends Biol. Sci. 21, 494 (1996).
    • (1996) Trends Biol. Sci. , vol.21 , pp. 494
    • Mann, M.1
  • 42
    • 1842330157 scopus 로고    scopus 로고
    • note
    • Several peptides were identical matches to human EST clones. All the EST clones were similar to human and mouse CHUK-1 (IKK-1). These clones were obtained (Genome Systems, Inc.) and the precise nucleotide sequence was determined and used to design primers to clone human IKK-2 by polymerase chain reaction (PCR) from a human HeLa cell cDNA library (Clontech, Inc.). Several IKK-2 cDNA clones were isolated and sequenced. Full-length mouse IKK-1 and a partial human IKK-1 nucleotide sequence were available in the comprehensive database. Primers were designed to clone by PCR the human and mouse IKK-1 cDNAs. The partial human IKK-1 coding region was used to probe a HeLa cDNA phage library (Stratagene) to obtain the full-length human IKK-1 cDNA clone by standard procedures.
  • 43
    • 1842404602 scopus 로고    scopus 로고
    • note
    • For in vitro translation studies, HA-tagged IKK-1 and Flag-tagged IKK-2 were in vitro translated in RRLs, either separately or alone, exactly as described in the manufacturer's protocol (Promega).
  • 45
    • 1842374898 scopus 로고    scopus 로고
    • note
    • For immunocytochemistry, HeLa cells were transiently transfected with either HA-tagged IKK-1 or Flagtagged IKK-2 as indicated. Cells were washed with PBS, fixed with 4% paraformaldehyde (Electron Microscopy Sciences) for 30 min, and permeabilized with wash buffer (0.5% Triton X-100, 0.01 % azide in PBS). Cells were blocked with wash buffer containing donkey serum (5%) (Jackson lmmunoresearch Laboratories) and probed with primary antibody-anti-RelA (polyclonal antibody), 1:2000 (Santa Cruz); anti-HA (monoclonal antibody), 1:1000 (Eastman Kodak); or anti-Flag (monoclonal antibody), 1:1000 (Babco)-followed by secondary antibody-donkey antibody to rabbit (fluorescein isothiocyanate) conjugate, 1:100; donkey antibody to mouse (Texas Red conjugate), 1:100 (Jackson Immunoresearch Laboratories). Cover slips were mounted with polyvinyl alcohol and 1,4-diazabicyclo[2.2.2]octane (Sigma) mounting medium and the slides were viewed under fluorescence with a Nikon Microphot-FXA microscope; the images were then scored and photographed.
  • 47
    • 0027295420 scopus 로고
    • A. Kumar, J. Haque, J. Lacoste, J. Hiscott, B. R. G. Williams, Proc. Natl. Acad. Sci. U.S.A. 91, 6288 (1994); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); M. T. Diaz-Meco et al., EMBO J. 13, 1842 (1994); G. J. Schouten et al., ibid. 16, 3133 (1997).
    • (1993) Mol. Cell. Biol. , vol.13 , pp. 4770
    • Diaz-Meco, M.T.1
  • 48
    • 85177153663 scopus 로고
    • A. Kumar, J. Haque, J. Lacoste, J. Hiscott, B. R. G. Williams, Proc. Natl. Acad. Sci. U.S.A. 91, 6288 (1994); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); M. T. Diaz-Meco et al., EMBO J. 13, 1842 (1994); G. J. Schouten et al., ibid. 16, 3133 (1997).
    • (1994) EMBO J. , vol.13 , pp. 1842
    • Diaz-Meco, M.T.1
  • 49
    • 0031007867 scopus 로고    scopus 로고
    • A. Kumar, J. Haque, J. Lacoste, J. Hiscott, B. R. G. Williams, Proc. Natl. Acad. Sci. U.S.A. 91, 6288 (1994); M. T. Diaz-Meco et al., Mol. Cell. Biol. 13, 4770 (1993); M. T. Diaz-Meco et al., EMBO J. 13, 1842 (1994); G. J. Schouten et al., ibid. 16, 3133 (1997).
    • (1997) EMBO J. , vol.16 , pp. 3133
    • Schouten, G.J.1
  • 51
    • 0031586174 scopus 로고    scopus 로고
    • J. A. DiDonato, M. Hayakawa, D. M. Rothwarf, E. Zandi, M. Karin, Nature 388, 548 (1997); C. H. Regnier et al., Cell 90, 373 (1997).
    • (1997) Cell , vol.90 , pp. 373
    • Regnier, C.H.1
  • 53
    • 1842413494 scopus 로고    scopus 로고
    • note
    • We thank I. Verma and R. Mueller for the gift of the IκB-α COOH-terminus construct and helpful discussions; J. DiDonato for the IκB-α vectors; H. Raymon and N. Richard for assistance with immunocytochemistry; S. Kim, A. LaPointe, and B. Lee for technical assistance; K. Davis and E. Carlson for help with compiling the manuscript and figures; and D. Anderson and A. Lewis for helpful comments and support. A.R. contributed to this work during a sabbatical at Signal Pharmaceuticals.


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