Sequence specific DNA binding by p53

A method of pre-screening agents for use in cancer therapy, comprising:

• contacting a transfected cell with a test substance, said transfected cell containing a p53 protein which is encoded by a mutant gene found in cancer cells of a patient and a reporter gene construct comprising a reporter gene which encodes an assayable product and a sequence which conforms to the p53 consensus binding site having more than one monomer of the double-stranded sequence RRRCWWGYYY, wherein said sequence is upstream from and adjacent to said reporter gene;
• and
• determining whether the amount of expression of said reporter gene is altered by the test substance, a test substance which alters the amount of expression of said reporter gene being a candidate for use in cancer therapy.

A method of pre-screening agents for use in cancer therapy, comprising:

• adding RNA polymerase and ribonucleotides to a transcription construct, said transcription construct comprising a reporter gene which encodes an assayable product and a sequence which conforms to the p53 consensus binding site having at least two monomers of the double-stranded sequence RRRCWWGYYY, said sequence being upstream from and adjacent to said reporter gene, said step of adding being effected in the presence and absence of a test substance;
• and
• determining whether the amount of transcription of said reporter gene is altered by the presence of said test substance, a test substance which alters the amount of transcription of said reporter gene being a candidate for use in cancer therapy.

A DNA construct for use in screening potential chemotherapeutic agents, comprising:

• a reporter gene which encodes an assayable product;
• a sequence which conforms to the p53 consensus binding site having more than one monomer of the double-stranded sequence RRRCWWGYYY upstream from and adjacent to said reporter gene, wherein said DNA construct is selected from the group consisting of a recombinant plasmid, a vital vector and an isolated molecule of DNA.

A method of pre-screening agents for use in cancer therapy, comprising:

• contacting a transfected cell with a test substance, said transfected cell containing a wild-type p53 protein and a reporter gene construct comprising a reporter gene which encodes an assayable product and a sequence which conforms to the p53 consensus binding site having more than one monomer of double-stranded sequence RRRCWWGYYY, wherein said sequence is upstream from and adjacent to said reporter gene;
• and
• determining whether the amount of expression of said reporter gene is altered by the test substance, a test substance which alters the amount of expression of said reporter gene being a candidate for use in cancer therapy.

A method of pre-screening oligonucleotides for use in cancer therapy, comprising:

• adding (a) a p53 protein which is encoded by a mutant gene found in a cancer patient and (b) a preparation of random oligonucleotides to (c) a p53-specific-binding DNA fragment immobilized on a solid support, to bind p53 protein to said p53-specific-binding DNA fragment immobilized on a solid support, wherein said p53-specific-binding DNA fragment contains more than one monomer of the double stranded motif RRRCWWGYYY;
• and
• recovering oligonucleotides from said preparation which bound to p53 which bound to said p53-specific-binding DNA fragment immobilized on the solid support.

A method of pre-screening agents for use in cancer therapy, comprising:

• contacting a transfected cell with a test substance, said transfected cell containing a p53 protein which is encoded by a mutant gene found in cancer cells of a patient and reporter gene construct comprising a reporter gene which encodes an assayable product and a sequence which conforms to the p53 consensus binding site having more than two monomers of TGCCT, wherein said sequence is upstream from and adjacent to said reporter gene;
• determining whether the amount of expression of said reporter gene is altered by the test substance, a test substance which alters the amount of expression of said reporter gene being a candidate for use in cancer therapy.

A method of pre-screening agents for use in cancer therapy, comprising:

• adding RNA polymerase and ribonucleotides to a transcription construct comprising a reporter gene which encodes an assayable enzyme activity and a sequence which conforms to the p53 consensus binding site having more than two monomers of TGCCT, said sequence being upstream from and adjacent to said reporter gene, said step of adding being effected in the presence and absence of the test substance;
• and
• determining whether the amount of transcription of said reporter gene is altered by the presence of said test substance, a test substance which alters the amount of transcription of said reporter gene being a candidate for use in cancer therapy.

A DNA construct for use in screening potential chemotherapeutic agents, comprising:

• a reporter gene which encodes an assayable product;
• a sequence which conforms to the p53 consensus binding site having more than two monomers of TGCCT upstream from and adjacent to said reporter gene, wherein said DNA construct is selected from the group consisting of a recombinant plasmid, a viral vector or an isolated molecule of DNA.

A method of pre-screening agents for use in cancer therapy, comprising:

• contacting a transfected cell with a test substance, said transfected cell containing a wild-type p53 protein and a reporter gene construct comprising a reporter gene which encodes an assayable product and a sequence which conforms to the p53 consensus binding site having more than two monomers of TGCCT, wherein said sequence is upstream from and adjacent to said reporter gene;
• and
• determining whether the amount of expression of said reporter gene is altered by the test substance, a test substance which alters the amount of expression of said reporter gene being a candidate for use in cancer therapy.

A method of pre-screening oligonucleotides for use in cancer therapy, comprising:

• adding (a) a p53 protein which is encoded by a mutant gene found in a cancer patient and (b) a preparation of random oligonucleotides to (c) a p53-specific-binding DNA fragment immobilized on a solid support, to bind p53 protein to said p53-specific-binding DNA fragment immobilized on a solid support, wherein said p53-specific-binding DNA fragment contains more than two monomers of the monomer sequence TGCCT;
• and
• recovering oligonucleotides from said preparation which bound to p53 which bound to said p53-specific-binding DNA fragment immobilized on the solid support.