INHIBITION OF NUCLEAR-PROTEIN BINDING TO THE HUMAN KI-RAS PROMOTER BY TRIPLER-FORMING OLIGONUCLEOTIDES

被引:38
作者
MAYFIELD, C [1 ]
SQUIBB, M [1 ]
MILLER, D [1 ]
机构
[1] UNIV ALABAMA, CTR COMPREHENS CANC, DEPT BIOCHEM, BIRMINGHAM, AL 35294 USA
关键词
D O I
10.1021/bi00177a029
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The human Ki-ras promoter contains a 22 base pair homopurine homopyrimidine (pur.pyr) motif within a region that is nuclease-hypersensitive in both native chromatin and supercoiled plasmids. Gel mobility shift analysis and competition experiments show that this pur.pyr motif binds a nuclear protein(s) in a sequence-specific manner. Several observations suggest that the nuclear protein may be an important regulatory factor. Gel mobility shift analysis and DNase I footprinting demonstrate that oligonucleotides can be targeted to this motif forming sequence-specific purine*purine.pyrimidine (pur*pur.pyr) or mixed purine/pyrimidine*purine.pyrimidine (pur/pyr*pur.pyr) intermolecular triple helices through guanine (G) recognition of guanine cytosine (G.C) base pairs and either adenine (A) or thymine (T) recognition of adenine thymine (A.T) base pairs in the target sequence. Triple helices containing either T*A.T or A*A.T triplets are formed exclusively with oligonucleotides antiparallel to the homopurine target strand. The affinity of an oligonucleotide which forms T*A.T triplets is approximately equal to, or slightly greater than, the affinity of an oligonucleotide which forms A*A.T triplets. Oligonucleotide-directed tripler formation inhibits sequence-specific nuclear protein binding to the K-ras promoter. These observations suggest that tripler formation by the oligonucleotides described here may provide a means to specifically inhibit transcription of the K-ras oncogene.
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页码:3358 / 3363
页数:6
相关论文
共 32 条
  • [1] MOST HUMAN CARCINOMAS OF THE EXOCRINE PANCREAS CONTAIN MUTANT C-K-RAS GENES
    ALMOGUERA, C
    SHIBATA, D
    FORRESTER, K
    MARTIN, J
    ARNHEIM, N
    PERUCHO, M
    [J]. CELL, 1988, 53 (04) : 549 - 554
  • [2] RAS GENES
    BARBACID, M
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1987, 56 : 779 - 827
  • [3] THE INFLUENCE OF SINGLE BASE TRIPLET CHANGES ON THE STABILITY OF A PUR-BULLET-PUR-BULLET-PYR TRIPLE HELIX DETERMINED BY AFFINITY CLEAVING
    BEAL, PA
    DERVAN, PB
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (11) : 2773 - 2776
  • [4] 2ND STRUCTURAL MOTIF FOR RECOGNITION OF DNA BY OLIGONUCLEOTIDE-DIRECTED TRIPLE-HELIX FORMATION
    BEAL, PA
    DERVAN, PB
    [J]. SCIENCE, 1991, 251 (4999) : 1360 - 1363
  • [5] THE MOLECULAR-GENETICS OF CANCER
    BISHOP, JM
    [J]. SCIENCE, 1987, 235 (4786) : 305 - 311
  • [6] PREVALENCE OF RAS GENE-MUTATIONS IN HUMAN COLORECTAL CANCERS
    BOS, JL
    FEARON, ER
    HAMILTON, SR
    VERLAANDEVRIES, M
    VANBOOM, JH
    VANDEREB, AJ
    VOGELSTEIN, B
    [J]. NATURE, 1987, 327 (6120) : 293 - 297
  • [7] BOS JL, 1989, CANCER RES, V49, P4682
  • [8] DIGNAM JD, 1990, METHOD ENZYMOL, V182, P194
  • [9] BINDING OF TRIPLE HELIX FORMING OLIGONUCLEOTIDES TO SITES IN GENE PROMOTERS
    DURLAND, RH
    KESSLER, DJ
    GUNNELL, S
    DUVIC, M
    PETTITT, BM
    HOGAN, ME
    [J]. BIOCHEMISTRY, 1991, 30 (38) : 9246 - 9255
  • [10] DURLAND RH, 1990, JERUS SYM Q, V23, P565