Claims for Patent: 5,908,762
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Summary for Patent: 5,908,762
Title: | Transcription factor regulating MHC expression CDNA and genomic clones encoding same and retroviral expression constructs thereof |
Abstract: | The present invention relates to NF-X1, a novel DNA binding protein which regulates expression of major histocompatibility complex (MHC) class II molecules, and to DNA sequences which encode the protein as well as recombinant expression of the protein. NF-X1 is a newly identified, cysteine-rich polypeptide which interacts sequence-specifically with the conserved X1 box regulatory element found in the proximal promoters of class II MHC genes. A cysteine-rich domain within NF-X1 contains a motif repeated seven times, and this entire region is necessary and sufficient for both sequence specific binding and effector function. The motif is related to but distinct from the previously described metal-binding protein families: LIM domain and RING finger. NFX.1 mRNA is markedly overexpressed late after induction of cells with interferon-gamma, and this overexpression coincides with a reduction in the level of HLA-DRA transcript in these cells. Overexpression of this protein strongly and specifically represses the transcription of the HLA-DRA gene in MHC class II positive cell lines, indicating that the NF-X1 protein is a transcriptional repressor of MHC class II molecules. |
Inventor(s): | Ono; Santa Jeremy (Baltimore, MD), Strominger; Jack L. (Lexington, MA) |
Assignee: | The Johns Hopkins University (Cambridge, MA) |
Application Number: | 08/828,584 |
Patent Claims: | 1. A chimeric DNA molecule comprising a DNA sequence that encodes a polypeptide having a sequence corresponding to SEQ ID NO:5, the sequence of said polypeptide selected from the group
consisting of SEQ ID NO:5, muteins of SEQ ID NO:5, truncations of SEQ ID NO:5, and fusion proteins containing them, wherein said polypeptide specifically binds to double stranded DNA having the sequence of SEQ ID NO:3.
2. The chimeric DNA molecule of claim 1, wherein said DNA sequence encoding said polypeptide is an intron-free DNA sequence. 3. A cell population transformed with the DNA molecule of claim 1, said population being substantially free of cells not transformed with said DNA molecule. 4. A composition comprising the chimeric DNA molecule of claim 1 and a pharmaceutically acceptable carrier. 5. A method of producing a polypeptide which contains at least about five copies of a cysteine rich sequence according to SEQ ID NO:1 coupled to a cysteine rich sequence according to SEQ ID NO:2 by a bridge peptide of from one to five amino acids, said polypeptide specifically binding to double stranded DNA having a sequence according to SEQ ID NO:3, comprising: providing a population of transformed cells of claim 3; growing said population under conditions whereby said polypeptide is expressed; and recovering said polypeptide. 6. The method of claim 5 where said polypeptide is excreted by said cell. 7. A DNA molecule comprising an intron-free DNA sequence encoding the amino acid sequence of SEQ ID NO:5. 8. The DNA molecule according to claim 7, corresponding to SEQ ID NO:4. 9. The DNA molecule according to claim 7 comprising a DNA sequence encoding NF-X1 found in a plasmid deposited under ATCC Accession No. 75895. 10. A single stranded DNA molecule comprising at least 20 sequential nucleotides, wherein said sequential nucleotides comprise a subsequence found in SEQ ID NO:4 or a DNA sequence complementary thereto. 11. A method for determining the presence of a polynucleotide substantially homologous to a coding sequence for a polypeptide which contains at least about five copies of a cysteine rich sequence according to SEQ ID NO:1 coupled to a cysteine rich sequence according to SEQ ID NO:2 by a bridge peptide of from one to five amino acids, said polypeptide specifically binding to double stranded DNA having a sequence according to SEQ ID NO:3, comprising: providing a sample suspected of comprising said polynucleotide; incubating the sample with a nucleotide probe having a sequence complementary to the single stranded DNA of claim 1, under conditions where said probe will form hybrids with nucleic acids substantially homologous to said probe, detecting nucleic acid hybrids; and the presence of nucleic acid hybrids indicating the presence of said polynucleotide. |
Details for Patent 5,908,762
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | 06/04/1986 | ⤷ Try a Trial | 2014-10-21 |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | ⤷ Try a Trial | 2014-10-21 | |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | Injection | 103132 | ⤷ Try a Trial | 2014-10-21 | |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
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