Tumor necrosis factor receptor-associated factors
AssigneeGenentech, Inc.
InventorsDavid V. Goeddel, Mike Rothe
Life of this patent
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Abstract
The invention concerns new tumor necrosis factor receptor associated factors, designated TRAF. The new factors are capable of specific association with the intracellular domain of the type 2 TNF receptor (TNF-R2), and are involved in the mediation of TNF biological activities.
Claims33
An isolated tumor necrosis receptor associated factor (TRAF) capable of specific association with the intracellular domain of a native type 2 TNF receptor (TNF-R2).
A molecule capable of disrupting the interaction of a TRAF and a native TNF-R2.
An antibody capable of specific binding to a native TRAF polypeptide.
A method of using a nucleic acid molecule encoding a TRAF comprising expressing such nucleic acid molecule in a cultured host cell transformed with a vector comprising such nucleic acid molecule operably linked to control sequences recognized by said host cell, and recovering the polypeptide encoded by said nucleic acid molecule from the host cell.
A method for producing a TRAF polypeptide comprising inserting into the DNA of a cell containing nucleic acid encoding said polypeptide a transcription modulatory element in sufficient proximity and orientation to the nucleic acid molecule to influence the transcription thereof.
A method of determining the presence of a TRAF polypeptide comprising hybridizing DNA encoding such polypeptide to a test sample nucleic acid and determining the presence of TRAF polypeptide DNA.
An isolated nucleic acid molecule encoding a fusion of an intracellular domain sequence of a native TNF-R2 to the DNA-binding domain of a transcriptional activator.
An isolated nucleic acid molecule encoding a fusion of a TRAF to the activation domain of a transcriptional activator.
An assay for identifying a factor capable of specific binding to the intracellular domain of a native TNF-R2, comprising
- (a) expressing nucleic acid molecules encoding a polypeptide comprising a fusion of an intracellular domain sequence of a native TNF-R2 to the DNA-binding domain of a transcriptional activator, and a fusion of a candidate polypeptide factor to the activation domain of a transcriptional activator, in a single host cell carrying a reporter gene;
- (b) monitoring the binding of said candidate factor to the intracellular domain of TNF-R2 by detecting a signal of the molecule encoded by said reporter gene.
As assay for identifying a factor capable of specific association with the intracellular domain of a native TNF-R2, comprising
- (a) expressing nucleic acid molecules encoding a polypeptide comprising a fusion of an intracellular domain sequence of a native TNF-R2 to the DNA-binding domain of a transcriptional activator, and a fusion of a candidate factor to the activation domain of a transcriptional activator, in a single host cell transfected with nucleic acid encoding a polypeptide factor capable of specific binding to said TNF-R2, and with nucleic acid encoding a reporter gene;
- and
- (b) monitoring the association of said candidate factor with said TNF-R2 or with said polypeptide factor capable of specific binding to said TNF-R2 by detecting the signal of the polypeptide encoded by said reporter gene.
A method of amplifying a nucleic acid test sample, comprising priming a nucleic acid polymerase reaction with nucleic acid encoding a TRAF polypeptide capable of specific association with the intracellular domain of a native TNF-R2.
A method for detecting a nucleic acid sequence coding for a polypeptide molecule which comprises all or part of a TRAF polypeptide or a related nucleic acid sequence, comprising contacting the nucleic acid sequence with a detectable marker which binds specifically to at least part of said nucleic acid sequence, and detecting the marker so bound.
A method for the prevention or treatment of a pathological condition associated with a TNF biological activity mediated, fully or partially, by a TNF-R2, comprising administering to a patient in need a preventatively or therapeutically effective amount of a TRAF or a molecule capable of disrupting the interaction of a TRAF and said TNF-R2.
An isolated human tumor necrosis factor receptor associated factor (TRAF) encoded by a DNA molecule obtainable from a human recombinant cDNA library or genomic DNA library (a) by the yeast two-hybrid technique or (b) by cross-species hybridization with an oligonucleotide sequence derived from the nucleotide sequence encoding murine TRAF1 (SEQ. ID. NO:
- 1) or murine TRAF2 (SEQ. ID. NO:
- 3).
An isolated human tumor necrosis factor receptor associated factor (TRAF) which is the human homolog of the murine TNFR1 (SEQ. ID. NO:
- 2).
An isolated human tumor necrosis factor receptor associated factor (TRAF) which is the human homolog of the murine TNFR2 (SEQ. ID. NO:
- 4).
Dependencies are inferred from the claim text.