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B. Kempkes et al., EMBO J. 14, 88 (1995). To construct the pTCMyc vector, we subcloned an SV40 polyadenylate [poly(A)] sequence and a Hind III genomic fragment containing exons 2 and 3 of c-MYC into the pTC vector [A. Polack et al., Proc. Natl. Acad. Sci. U.S.A. 93, 10411 (1996)].
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Kempkes, B.1
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B. Kempkes et al., EMBO J. 14, 88 (1995). To construct the pTCMyc vector, we subcloned an SV40 polyadenylate [poly(A)] sequence and a Hind III genomic fragment containing exons 2 and 3 of c-MYC into the pTC vector [A. Polack et al., Proc. Natl. Acad. Sci. U.S.A. 93, 10411 (1996)].
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Polack, A.1
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0344072456
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K.-J. Wu, A. Polack, R. Dalla-Favera, data not shown
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K.-J. Wu, A. Polack, R. Dalla-Favera, data not shown.
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19
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0344503707
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note
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1, after withdrawal of E2, and this was not changed by c-MYC induction after TC withdrawal.
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20
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T. D. Littlewood, D. C. Hancock, P. C. Danielian, M. G. Parker, G. I. Evan, Nucleic Acids Res. 23, 1686 (1995).
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M. A. Bevilacqua et al., Biochem. J. 311, 769 (1995); M. W. Hentze et al., Proc. Natl. Acad. Sci. U.S.A. 83, 7226 (1986). A polymerase chain reaction (PCR) fragment containing the H-ferritin promoter (nucleotides -160 to +36) was subcloned into a pXP2 promoterless luciferase vector to generate the FPLuc vector. The FPMutLuc vector was generated by site-directed mutagenesis.
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M. A. Bevilacqua et al., Biochem. J. 311, 769 (1995); M. W. Hentze et al., Proc. Natl. Acad. Sci. U.S.A. 83, 7226 (1986). A polymerase chain reaction (PCR) fragment containing the H-ferritin promoter (nucleotides -160 to +36) was subcloned into a pXP2 promoterless luciferase vector to generate the FPLuc vector. The FPMutLuc vector was generated by site-directed mutagenesis.
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(1986)
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Hentze, M.W.1
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The pMT2TMyc expression vector was constructed by subcloning a fragment containing exons 2 and 3 of MYC genomic DNA into the Eco RI site of the pMT2T vector; pMT2TMycΔMBII and pMT2TMycΔHLH were made by substituting in pMT2TMyc exons 2 and 3 of c-MYC from the SVSP65MLVMyc vector [J. Stone et al., Mol. Cell. Biol. 7, 1697 (1987)], which contains deletions of amino acids 122 to 140 and 371 to 412, respectively.
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Stone, J.1
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0344072455
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Supplementary data
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Supplementary data are available at www.sciencemag. org/feature/data/983675.shl.
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R. D. Klausner, T. A. Rouault, J. B. Harford, Cell 72, 19 (1993); M. W. Hentze and L. C. Kuhn, Proc. Natl. Acad. Sci. U.S.A. 93, 8175 (1996).
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R. D. Klausner, T. A. Rouault, J. B. Harford, Cell 72, 19 (1993); M. W. Hentze and L. C. Kuhn, Proc. Natl. Acad. Sci. U.S.A. 93, 8175 (1996).
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Iwai, K.1
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0344934488
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note
-
The pTCFer expression vector was constructed by subdoning an H-ferritin cDNA fragment containing the complete coding domain (+86 to +665) (15) into the Not I site of pTC-SV.
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C. D. Gregory et al., Nature 349, 612 (1991).
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0344072453
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note
-
The pHeBoCMVFerritin expression vector was made by subcloning an H-ferritin cDNA fragment into the Nhe I-Not I sites of pHeBoCMV vector (4).
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35
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0026013466
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H. E. Broxmeyer, S. Cooper, S. Levi, P. Arosio, Proc. Natl. Acad. Sci. U.S.A. 88, 770 (1991). Treatment with iron-free hemin (protoporphyrin IX) led to no colony formation for all cell types [E. Coccia et al., Eur. J. Biochem. 250, 764 (1997)].
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Broxmeyer, H.E.1
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H. E. Broxmeyer, S. Cooper, S. Levi, P. Arosio, Proc. Natl. Acad. Sci. U.S.A. 88, 770 (1991). Treatment with iron-free hemin (protoporphyrin IX) led to no colony formation for all cell types [E. Coccia et al., Eur. J. Biochem. 250, 764 (1997)].
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S. J. Elledge, Z. Zhou, J. B. Allen, Trends Biochem. Sci. 17, 119 (1992); C. E. Cooper et al., J. Biol. Chem. 271, 20291 (1996).
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S. J. Elledge, Z. Zhou, J. B. Allen, Trends Biochem. Sci. 17, 119 (1992); C. E. Cooper et al., J. Biol. Chem. 271, 20291 (1996).
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C. Brodie et al., Cancer Res. 53, 3968 (1993); K. P. Hoyes, R. C. Hider, J. B. Porter, ibid. 52, 4591 (1992); K. S. Kulp and P. R. Vulliet, Toxicol. Appl. Pharmacol. 139, 356 (1996).
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0011765668
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The H-ferritin probe was generated by reverse transcriptase (RT)-PCR amplification of U937 RNA. For the c-MYC exons 1 and 3, probes were obtained from the pMC41RC and pMC41ER plasmids [R. Dalla-Favera et al., Proc. Natl. Acad. Sci. U.S.A. 79, 6497 (1982)]. The SV40 probe was obtained by digesting the pHeBoCMVFerritin vector with the Bam HI and Sal I enzymes to generate a 400-base pair fragment. The 12S ribosomal RNA probe was obtained by RT-PCR from U937 RNA.
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(1982)
Proc. Natl. Acad. Sci. U.S.A.
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Dalla-Favera, R.1
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43
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0027198736
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4, 8 mM EDTA, and 2 mM phenylmethylsulfonyl fluoride) with a polyclonal antiserum to ferritins (Boehringer Mannheim) for 2 hours using protein A Sepharose. Immunoprecipitates were electrophoresed on a 12% SDS-PAGE minigel, transferred to nitrocellulose, and then incubated with a monoclonal antibody to human H-ferritin [P. Santambrogio et al., J. Biol. Chem. 268, 12744 (1993)]. Signal detection was carried out with sheep antibody to mouse immunoglobulins and an ECL kit (Amersham).
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J. Biol. Chem.
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Santambrogio, P.1
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44
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0345365892
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note
-
The human and rat IRP2 probes were generated by RT-PCR amplification of mRNAs from human lymphoblastoid cells and Rat1 cells, respectively. The primers used for the generation of the IRP2 probe were 5′-CCATAGCAGGCACAGTGAAT and 3′-GGAATCTGCATTTTCTCCTG (both identical for human and rat).
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45
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0029034338
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B. Guo, F. M. Brown, J. D. Philips, Y. Yu, E. A. Leibold, J. Biol. Chem. 270, 16529 (1995).
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Guo, B.1
Brown, F.M.2
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Yu, Y.4
Leibold, E.A.5
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46
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0345365890
-
-
note
-
For soft agar assays (4), three different cell numbers were plated in top agar in triplicate, and colonies were counted after 14 days of incubation at 37°C.
-
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-
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47
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0344503700
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note
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We thank B. Vogelstein, R. Eisenman, T. Littlewood, F. Costanzo, G. Bornkamm, P. Arosio, J. Sedivy, T. McGraw, E. Ziff, R. Liem, and E. Leibold for providing various reagents and cell lines and E. Marcantonio, B. Tycko, and A. Migliazza for critical reading of the manuscript. K.J.W. is a Fellow of the Leukemia Society of America. Supported by NIH grant CA-37165.
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