Regulation of gene expression with pyrrole-imidazole polyamides

Journal of Biotechnology

Volumn 112, Issue 1-2, 2004, Pages 195-220

  Melander, Christian ^a   Burnett, Ryan ^a   Gottesfeld, Joel M ^a  

^a SCRIPPS RESEARCH INSTITUTE   (United States)

Author keywords

DNA minor groove; Gene regulation; Pyrrole imidazole polyamide; Transcription

Indexed keywords

CELL CULTURE; CELLS; DNA; GENES; GENETIC ENGINEERING; MATHEMATICAL MODELS; PROTEINS;

EUKARYOTIC TRANSCRIPTIONAL REGULATORY PROTEINS; GENE EXPRESSIONS; POTENT INHIBITORS; PYRROLE-IMIDAZOLE (PY-IM) POLYAMIDES;

POLYAMIDES;

CONGOCIDINE; DISTAMYCIN A; DNA BINDING PROTEIN; DOUBLE STRANDED DNA; ESTROGEN RECEPTOR; IMIDAZOLE DERIVATIVE; LYMPHOID ENHANCER FACTOR 1; MESSENGER RNA; POLYAMIDE; PYRROLE DERIVATIVE; REGULATOR PROTEIN; TRANSCRIPTION FACTOR ETS 1;

ANTIFUNGAL ACTIVITY; ARTICLE; BINDING AFFINITY; BINDING SITE; CELL CULTURE; CELL LINE; CELL MEMBRANE PERMEABILITY; CELLULAR DISTRIBUTION; CHROMATIN; CHROMOSOME; CRYSTAL STRUCTURE; DNA BINDING; DNA SEQUENCE; DNA TEMPLATE; DOWN REGULATION; DROSOPHILA; GENE ACTIVATION; GENE CONTROL; GENE EXPRESSION; GENE TARGETING; GENOME; HUMAN; HUMAN IMMUNODEFICIENCY VIRUS 1; LIGAND BINDING; MOLECULAR RECOGNITION; NONHUMAN; PRIORITY JOURNAL; PROTEIN DNA INTERACTION; PROTEIN FAMILY; T LYMPHOCYTE; TATA BOX; VIRUS REPLICATION; VIRUS TRANSCRIPTION;

ANIMALS; DNA; DNA-BINDING PROTEINS; DROSOPHILA MELANOGASTER; GENE EXPRESSION REGULATION; HUMANS; IMIDAZOLES; NYLONS; PYRROLES; SIGNAL TRANSDUCTION;

EUKARYOTA;

EID: 3843142798     PISSN: 01681656     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.jbiotec.2004.03.018     Document Type: Article

References (103)

1. Ansari A.Z., Mapp A.K., Nguyen D.H., Dervan P.B., Ptashne M. Towards a minimal motif for artificial transcriptional activators. Chem. Biol. 8:2001;583-592
2. Arora P.S., Ansari A.Z., Best T.P., Ptashne M., Dervan P.B. Design of artificial transcriptional activators with rigid poly-L-proline linkers. J. Am. Chem. Soc. 124:2002;13067-13071
3. Baird E.E., Dervan P.B. Solid phase synthesis of polyamides containing imidazole and pyrrole amino acids. J. Am. Chem. Soc. 118:1996;6141-6146
4. Bassuk A.G., Anandappa R.T., Leiden J.M. Physical interactions between Ets and NF-kappaB/NFAT proteins play an important role in their cooperative activation of the human immunodeficiency virus enhancer in T cells. J. Virol. 71:1997;3563-3573
5. Belitsky J.M., Leslie S.J., Arora P.S., Beerman T.A., Dervan P.B. Cellular uptake of N-methylpyrrole/N-methylimidazole polyamide-dye conjugates. Bioorg. Med. Chem. 10:2002;3313-3318
6. Best T.P., Edelson B.S., Nichols N.G., Dervan P.B. Nuclear localization of pyrrole-imidazole polyamide-fluorescein conjugates in cell culture. Proc. Natl. Acad. Sci. USA. 100:2003;12063-12068
7. Bremer R.E., Baird E.E., Dervan P.B. Inhibition of major-groove-binding proteins by pyrrole-imidazole polyamides with an Arg-Pro-Arg positive patch. Chem. Biol. 5:1998;119-133
8. Bremer R.E., Wurtz N.R., Szewczyk J.W., Dervan P.B. Inhibition of major groove DNA binding bZIP proteins by positive patch polyamides. Bioorg. Med. Chem. 9:2001;2093-2103
9. Chang A.Y., Dervan P.B. Strand selective cleavage of DNA by diastereomers of hairpin polyamide-seco-CBI conjugate. J. Am. Chem. Soc. 122:2000;4856-4864
10. Chen F.E., Huang D.-B., Chen Y.-Q., Ghosh G. Crystal structure of p50/p65 heterodimer of transcription factor NF-kB bound to DNA. Nature. 391:1998;410-413
11. Chiang S.Y., Burli R.W., Benz C.C., Gawron L., Scott G.K., Dervan P.B., Beerman T.A. Targeting the ets binding site of the HER2/neu promoter with pyrrole-imidazole polyamides. J. Biol. Chem. 275:2000;24246-24254
12. Choo Y., Sanchez-Garcia I., Klug A. In vivo repression by a site-specific DNA-binding protein designed against an oncogenic sequence. Nature. 372:1994;642-645
13. Coull J.J., He G., Melander C., Rucker V.C., Dervan P.B., Margolis D.M. Targeted derepression of the human immunodeficiency virus type 1 long terminal repeat by pyrrole-imidazole polyamides. J. Virol. 76:2002;12349-12354
14. Crowley K.S., Phillion D.P., Woodard S.S., Schweitzer B.A., Singh M., Shabany H., Burnette B., Hippenmeyer P., Heitmeier M., Bashkin J.K. Controlling the intracellular localization of fluorescent polyamide analogues in cultured cells. Bioorg. Med. Chem. Lett. 13:2003;1565-1570
15. Dervan P.B. Design of sequence-specific DNA-binding molecules. Science. 232:1986;464-471
16. Dervan P.B. Molecular recognition of DNA by small molecules. Bioorg. Med. Chem. 9:2001;2215-2235
17. Dickinson L.A., Gulizia R.J., Trauger J.W., Baird E.E., Mosier D.E., Gottesfeld J.M., Dervan P.B. Inhibition of RNA polymerase II transcription in human cells by synthetic DNA-binding ligands. Proc. Natl. Acad. Sci. USA. 95:1998;12890-12895
18. Dickinson L.A., Trauger J.W., Baird E.E., Dervan P.B., Graves B.J., Gottesfeld J.M. Inhibition of Ets-1 DNA binding and ternary complex formation between Ets-1, NF-kB, and DNA by a designed DNA-binding ligand. J. Biol. Chem. 274:1999a;12765-12773
19. Dickinson L.A., Trauger J.W., Baird E.E., Ghazal P., Dervan P.B., Gottesfeld J.M. Anti-repression of RNA polymerase II transcription by pyrrole-imidazole polyamides. Biochemistry. 38:1999b;10801-10807
20. Dudouet B., Burnett R., Dickinson L.A., Wood M.R., Melander C., Belitsky J.M., Edelson B., Wurtz N., Briehn C., Dervan P.B., Gottesfeld J.M. Accessibility of nuclear chromatin by DNA binding polyamides. Chem. Biol. 10:2003;859-867
21. Ehley J.A., Melander C., Herman D., Baird E.E., Ferguson H.A., Goodrich J.A., Dervan P.B., Gottesfeld J.M. Promoter scanning for transcription inhibition with DNA-binding polyamides. Mol. Cell. Biol. 22:2002;1723-1733
22. Fechter E.J., Dervan P.B. Allosteric inhibition of protein-DNA complexes by polyamide-intercalator conjugates. J. Am. Chem. Soc. 125:2003;8476-8485
23. Ferré-D'Amaré A.R., Prendergast G.C., Ziff E.B., Burley S.K. Recognition by Max of its cognate DNA through a dimeric b/HLH/Z domain. Nature. 363:1993;38-45
24. Foister S., Marques M.A., Doss R.M., Dervan P.B. Shape selective recognition of T.A base pairs by hairpin polyamides containing N-terminal 3-methoxy (and 3-chloro) thiophene residues. Bioorg. Med. Chem. 11:2003;4333-4340
25. Gearhart M.D., Holmbeck S.M., Evans R.M., Dyson H.J., Wright P.E. Monomeric complex of human orphan estrogen related receptor-2 with DNA: a pseudo-dimer interface mediates extended half-site recognition. J. Mol. Biol. 327:2003;819-832
26. Giese K., Pagel J., Grosschedl R. Functional analysis of DNA bending and unwinding by the high mobility group domain of LEF-1. Proc. Natl. Acad. Sci. USA. 94:1997;12845-12850
27. Gottesfeld J.M., Belitsky J.M., Melander C., Dervan P.B., Luger K. Blocking transcription through a nucleosome with synthetic DNA ligands. J. Mol. Biol. 321:2002;249-263
28. Gottesfeld J.M., Melander C., Suto R.K., Raviol H., Luger K., Dervan P.B. Sequence-specific recognition of DNA in the nucleosome by pyrrole-imidazole polyamides. J. Mol. Biol. 309:2001;615-629
29. Gottesfeld J.M., Neely L., Trauger J.W., Baird E.E., Dervan P.B. Regulation of gene expression by small molecules. Nature (London). 387:1997;202-205
30. Gottesfeld J.M., Turner J.M., Dervan P.B. Chemical approaches to control gene expression. Gene Exp. 9:2000;77-91
31. Graves B.J., Petersen J.M. Specificity within the ets family of transcription factors. Adv. Cancer Res. 75:1998;1-55
32. Gygi M.P., Ferguson M.D., Mefford H.C., Lund K.P., O'Day C., Zhou P., Friedman C., van den Engh G., Stolowitz M.L., Trask B.J. Use of fluorescent sequence-specific polyamides to discriminate human chromosomes by microscopy and flow cytometry. Nucleic Acids Res. 30:2002;2790-2799
33. Hannon G.J. RNA interference. Nature. 418:2002;244-251
34. He G., Margolis D.M. Counterregulation of chromatin deacetylation and histone deacetylase occupancy at the integrated promoter of human immunodeficiency virus type 1 (HIV-1) by the HIV-1 repressor YY1 and HIV-1 activator Tat. Mol. Cell. Biol. 22:2002;2965-2973
35. Henikoff S., Vermaak D. Bugs on drugs go GAGAA. Cell. 103:2000;695-698
36. Herman D.M., Baird E.E., Dervan P.B. Stereochemical control of the DNA binding affinity, sequence specificity, and orientation preference of chiral hairpin polyamides in the minor groove. J. Am. Chem. Soc. 120:1998;1382-1391
37. Herman D.M., Baird E.E., Dervan P.B. Tandem hairpin motif for recognition in the minor groove of DNA by pyrrole-imidazole polyamides. Chem. Eur. J. 5:1999a;975-983
38. Herman D.M., Turner J.M., Baird E.E., Dervan P.B. Cycle polyamide motif for recognition of the minor groove of DNA. J. Am. Chem. Soc. 121:1999b;1121-1129
39. Hernandez N. TBP, a universal eukaryotic transcription factor? Genes Dev. 7:1993;1291-1308
40. Janssen S., Cuvier O., Müller M., Laemmli U.K. Specific gain- and loss-of-function phenotypes induced by satellite-specific DNA-binding drugs fed to Drosophila melanogaster. Mol. Cell. 6:2000a;1013-1024
41. Janssen S., Durussel T., Laemmli U.K. Chromatin opening of DNA satellites by targeted sequence-specific drugs. Mol. Cell. 6:2000b;999-1011
42. Jones K.A., Peterlin B.M. Control of RNA initiation and elongation at the HIV-1 promoter. Annu. Rev. Biochem. 63:1994;717-743
43. Kielkopf C.L., Baird E.E., Dervan P.B., Rees D.C. Structural basis for GC recognition in the DNA minor groove. Nature Struct. Biol. 5:1998a;104-109
44. Kielkopf C.L., Bremer R.E., White S., Szewczyk J.W., Turner J.M., Baird E.E., Dervan P.B., Rees D.C. Structural effects of DNA sequence on TA recognition by hydroxypyrrole/pyrrole pairs in the minor groove. J. Mol. Biol. 295:2000;557-567
45. Kielkopf C.L., White S., Szewczyk J.W., Turner J.M., Baird E.E., Dervan P.B., Rees D.C. A structural basis for recognition of AT and TA base pairs in the minor groove of B-DNA. Science (Washington DC). 282:1998b;111-115
46. Kim J.L., Nikolov D.B., Burley S.K. Co-crystal structure of TBP recognizing the minor groove of a TATA element. Nature. 365:1993a;520-527
47. Kim Y., Geiger J.H., Hahn S., Sigler P.B. Crystal structure of a yeast TBP/TATA-box complex. Nature. 365:1993b;512-520
48. Kodandapani R., Pio F., Ni C.-Z., Piccialli G., Klemsz M., McKercher S., Maki R.A., Ely K.R. A new pattern for helix-turn-helix recognition revealed by the PU.1 ETS-domain-DNA complex. Nature. 380:1996;456-460
49. Kopka M.L., Yoon C., Goodsell D., Pjura P., Dickerson R.E. The molecular origin of DNA-drug specificity in netropsin and distamycin. Proc. Natl. Acad. Sci. USA. 82:1985;1376-1380
50. Lamamie de Clairac R.P., Geierstanger B.H., Mrksich M., Dervan P.B., Wemmer D.E. NMR characterization of hairpin polyamide complexes with the minor groove of DNA. J. Am. Chem. Soc. 119:1997;7909-7916
51. Lemon B., Tjian R. Orchestrated response: a symphony of transcription factors for gene control. Genes Dev. 14:2000;2551-2569
52. Lenzmeier B.A., Baird E.E., Dervan P.B., Nyborg J.K. The Tax protein-DNA interaction is essential for HTLV-I transactivation in vitro. J. Mol. Biol. 291:1999;731-744
53. Love J.J., Li X., Case D.A., Giese K., Grosschedl R., Wright P.E. Structural basis for DNA bending by the architectural transcription factor LEF-1. Nature. 376:1995;791-795
54. Luger K., Mader A.W., Richmond R.K., Sargent D.F., Richmond T.J. Crystal structure of the nucleosome core particle at 2.8 Å resolution. Nature. 389:1997;251-260
55. Maeshima K., Janssen S., Laemmli U.K. Specific targeting of insect and vertebrate telomeres with pyrrole and imidazole polyamides. EMBO J. 20:2001;3218-3228
56. Mapp A.K., Ansari A.Z., Ptashne M., Dervan P.B. Activation of gene expression by small molecule transcription factors. Proc. Natl. Acad. Sci. USA. 97:2000;3930-3935
57. Marini N.J., Baliga R., Taylor M.J., White S., Simpson P., Tsai L., Baird E.E. DNA binding hairpin polyamides with antifungal activity. Chem. Biol. 10:2003;635-644
58. McBryant S.J., Baird E.E., Trauger J.W., Dervan P.B., Gottesfeld J.M. Minor groove DNA-protein contacts upstream of a tRNA gene detected with a synthetic DNA binding ligand. J. Mol. Biol. 286:1999;973-981
59. Mrksich M., Dervan P.B. Design of a covalent peptide heterodimer for sequence-specific recognition in the minor groove of double-helical DNA. J. Am. Chem. Soc. 116:1994;3663-3664
60. Mrksich M., Parks M.E., Dervan P.B. Hairpin peptide motif. A new class of oligopeptides for sequence-specific recognition in the minor groove of double-helical DNA. J. Am. Chem. Soc. 116:1994;7983-7988
61. Mrksich M., Wade W.S., Dwyer T.J., Geierstanger B.H., Wemmer D.E., Dervan P.B. Antiparallel side-by-side dimeric motif for sequence-specific recognition in the minor groove of DNA by the designed peptide 1-methylimidazole-2- carboxamide netropsin. Proc. Natl. Acad. Sci. USA. 89:1992;7586-7590
62. Neely L., Trauger J.W., Baird E.E., Dervan P.B., Gottesfeld J.M. Importance of minor groove binding zinc fingers within the transcription factor IIIA DNA complex. J. Mol. Biol. 274:1997;439-445
63. Nevado J., Gaudreau L., Adam M., Ptashne M. Transcriptional activation by artificial recruitment in mammalian cells. Proc. Natl. Acad. Sci. USA. 96:1999;2674-2677
64. Nolte R.T., Conlin R.M., Harrison S.C., Brown R.S. Differing roles for zinc fingers in DNA recognition: structure of a six-finger transcription factor IIIA complex. Proc. Natl. Acad. Sci. USA. 95:1998;2938-2943
65. Oakley M.G., Mrksich M., Dervan P.B. Evidence that a minor groove-binding peptide and a major groove-binding protein can simultaneously occupy a common site on DNA. Biochemistry. 31:1992;10969-10975
66. Orphanides G., Lagrange T., Reinberg D. The general transcription factors of RNA polymerase II. Genes Dev. 10:1996;2657-2683
67. Pelton J.G., Wemmer D.E. Structural characterization of a 2-1 distamycin A-d(CGCAAATTTGGC) complex by two-dimensional. NMR. 86:1989;5723-5727
68. Rastinejad F., Perlmann T., Evans R.M., Sigler P.B. Structural determinants of nuclear receptor assembly on DNA direct repeats. Nature. 375:1995;203-211
69. Renneberg D., Dervan P.B. Imidazopyridine/Pyrrole and hydroxybenzimidazole/pyrrole pairs for DNA minor groove recognition. J. Am. Chem. Soc. 125:2003;5707-5716
70. Schaal T.D., Mallet W.G., McMinn D.L., Nguyen N.V., Sopko M.M., John S., Parekh B.S. Inhibition of human papilloma virus E2 DNA binding protein by covalently linked polyamides. Nucleic Acids Res. 31:2003;1282-1291
71. Schultz P.G., Taylor J.S., Dervan P.B. Design synthesis of a sequence-specific DNA cleaving molecule. (Distamycin-EDTA)iron(II). J. Am. Chem. Soc. 104:1982;6861-6863
72. Schwabe J.W., Chapman L., Finch J.T., Rhodes D. The crystal structure of the estrogen receptor DNA-binding domain bound to DNA: how receptors discriminate between their response elements. Cell. 75:1993;567-578
73. Seeman N.C., Rosenberg J.M., Rich A. Sequence-specific recognition of double helical nucleic acids by proteins. Proc. Natl. Acad. Sci. USA. 73:1976;804-808
74. Sun X.-H., Baltimore D. An inhibitory domain of E12 transcription factor prevents DNA binding in E12 homodimers but not in E12 heterodimers. Cell. 64:1991;459-470
75. Supekova L., Pezacki J.P., Su A.I., Loweth C.J., Riedl R., Geierstanger B., Schultz P.G., Wemmer D.E. Genomic effects of polyamide/DNA interactions on mRNA expression. Chem. Biol. 9:2002;821-827
76. Suto R.K., Clarkson M.J., Tremethick D.J., Luger K. Crystal structure of a nucleosome core particle containing the variant histone H2A.Z. Nature Struct. Biol. 7:2000;1121-1124
77. Suto R.K., Edayathumangalam R.S., White C.L., Melander C., Gottesfeld J.M., Dervan P.B., Luger K. Effects of ligand binding on structure and dynamics of the nucleosome core particle. J. Mol. Biol. 326:2003;371-380
78. Taylor J.S., Schultz P.G., Dervan P.B. DNA affinity cleaving. Sequence specific cleavage of DNA by distamycin-EDTA.cntdot.iron(II) EDTA-distamycin. iron(II). Tetrahedron. 40:1984;457-465
79. Trauger J.W., Baird E.E., Dervan P.B. Cooperative hairpin dimers for recognition of DNA by Py-Im polyamides. Angew. Chemie. Int. Ed. 37:1998a;1421-1423
80. Trauger J.W., Baird E.E., Dervan P.B. Recognition of 16 base pairs in the minor groove of DNA by a pyrrole-imidazole polyamide dimer. J. Am. Chem. Soc. 120:1998b;3534-3535
81. Trauger J.W., Baird E.E., Dervan P.B. Subnanomolar sequence-specific recognition in the minor groove of DNA by designed ligands. Nature (London). 382:1996;559-561
82. Travis A., Amsterdam A., Belanger C., Grosschedl R. LEF-1, a gene encoding a lymphoid-specific protein with an HMG domain regulates T-cell receptor a enhancer function. Genes Dev. 5:1991;880-894
83. Turner J.M., Biard E.E., Dervan P.B. Recognition of 7 base-pair sequences in the minor groove of DNA by 10-ring pyrrole-imidazole polyamide hairpins. J. Am. Chem. Soc. 119:1997;7636-7644
84. Turner J.M., Swalley S.E., Baird E.E., Dervan P.B. Aliphatic/aromatic amino acid pairings for polyamide recognition in the minor groove of DNA. J. Am. Chem. Soc. 120:1998;6219-6226
85. Tutter A., Jones K.A. Chemicals that footprint DNA: hitting HIV-1 in the minor groove. Proc. Natl. Acad. Sci. USA. 95:1998;12739-12741
86. Urbach A.R., Dervan P.B. Toward rules for 1:1 polyamide:DNA recognition. Proc. Natl. Acad. Sci. USA. 98:2001;4343-4348
87. Wade W.S., Dervan P.B. Alteration of the sequence specificity of distamycin on DNA by replacement of an N-methylpyrrolecarboxamide with pyridine-2-carboxamide. J. Am. Chem. Soc. 109:1987;1574-1575
88. Wade W.S., Mrksich M., Dervan P.B. Design of peptides that bind in the minor groove of DNA at 5′-(A,T)G(A,T)C(A,T)-3′ sequences by a dimeric side by side motif. J. Am. Chem. Soc. 114:1992;8784-8794
89. Wang Y.D., Dziegielewski J., Chang A.Y., Dervan P.B., Beerman T.A. Cell-free and cellular activities of a DNA sequence selective hairpin polyamide-CBI conjugate. J. Biol. Chem. 277:2002;42431-42437
90. Wang Y.D., Dziegielewski J., Wurtz N.R., Dziegielewska B., Dervan P.B., Beerman T.A. DNA crosslinking and biological activity of a hairpin polyamide-chlorambucil conjugate. Nucleic Acids Res. 31:2003;1208-1215
91. Waterman M.L., Jones K.A. Purification of TCF-1a, a T-cell-specific transcription factor that activates the T-cell receptor gene enhancer in a context-dependent manner. New Biol. 2:1990;621-636
92. Wemmer D.E., Dervan P.B. Targeting the minor groove of DNA. Curr. Opin. Struct. Biol. 7:1997;355-361
93. White C.L., Suto R.K., Luger K. Structure of the yeast nucleosome core particle reveals fundamental changes in internucleosome interactions. EMBO J. 20:2001;5207-5218
94. White S., Szewczyk J.W., Turner J.M., Baird E.E., Dervan P.B. Recognition of the four Watson-Crick base pairs in the DNA minor groove by synthetic ligands. Nature (London). 391:1998;468-471
95. White S., Turner J.M., Szewczyk J.W., Baird E.E., Dervan P.B. Affinity and specificity of multiple hydroxypyrrole/pyrrole ring pairings for coded recognition of DNA. J. Am. Chem. Soc. 121:1999;260-261
96. Winston R.L., Ehley J.A., Baird E.E., Dervan P.B., Gottesfeld J.M. Asymmetric DNA binding by a homodimeric bHLH protein. Biochemistry. 39:2000;9092-9098
97. Winston R.L., Gottesfeld J.M. Rapid identification of key amino acid-DNA contacts through combinatorial peptide synthesis. Chem. Biol. 7:2000;245-251
98. Wolberger C. Multiprotein-DNA complexes in transcriptional regulation. Annu. Rev. Biophys. Biomol. Struct. 28:1999;29-56
99. Wurtz N.R., Dervan P.B. Sequence specific alkylation of DNA by hairpin pyrrole-imidazole polyamide conjugates. Chem. Biol. 7:2000;153-161
100. Wurtz N.R., Pomerantz J.L., Baltimore D., Dervan P.B. Inhibition of DNA binding by NF-kappa B with pyrrole-imidazole polyamides. Biochemistry. 41:2002;7604-7609
101. Youngquist R.S., Dervan P.B. A synthetic peptide binds 16 base pairs of A,T double helical DNA. J. Am. Chem. Soc. 109:1987;7564-7566
102. Zenzie-Gregory B., Sheridan P., Jones K.A., Smale S.T. HIV-1 core promoter lacks a simple initiator element but contains a bipartite activator at the transcription start site. J. Biol. Chem. 268:1993;15823-15832
103. Zhao Q., Khorasanizadeh S., Miyoshi Y., Lazar M.A., Rastinejad F. Structural elements of an orphan nuclear receptor-DNA complex. Mol. Cell. 1:1998;849-861