Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
1998-08-19
2002-07-16
McKelvey, Terry (Department: 1636)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Reexamination Certificate
active
06420525
ABSTRACT:
BACKGROUND OF THE INVENTION
Proliferation and differentiation of cells of multicellular organisms are controlled by hormones and polypeptide growth factors. These diffusable molecules allow cells to communicate with each other and act in concert to form cells and organs, and to repair and regenerate damaged tissue. Examples of hormones and growth factors include the steroid hormones (e.g. estrogen, testosterone), parathyroid hormone, follicle stimulating hormone, the interleukins, platelet derived growth factor (PDGF), epidermal growth factor (EGF), granulocyte-macrophage colony stimulating factor (GM-CSF), erythropoietin (EPO) and calcitonin.
Hormones and growth factors influence cellular metabolism by binding to receptors. Receptors may be integral membrane proteins that are linked to signaling pathways within the cell, such as second messenger systems. Other classes of receptors are soluble molecules, such as the transcription factors.
Transcription factors regulate the transcription of genes in the cell by interacting with other transcription factors and RNA polymerase. Transcription factors are characterized by their DNA-binding domain and their transcriptional activation domain. Within the DNA binding domain, several different motifs have been identified which act to mediate DNA binding to transcription factors. These include cysteine-histidine zinc finger and multi-cysteine zinc finger motifs, homeobox motifs, winged helix motifs, leucine-zipper motifs, and helix-loop-helix motifs. The activation domain can contain a large number of acetic amino acids which form an amphipathic &agr;-helix with its negative charges displayed on one surface. Others have glutamine or proline-rich regions.
Following DNA binding, the transcription factor interacts with other factors or the RNA polymerase to stimulate transcription. Transcription can be blocked by molecules which are able to bind to the DNA binding domain but do not interact with the transcription domain. These repressor molecules prevent positively acting DNA molecules from binding.
The location of the transcription factors bound by a particular gene control the gene's expression pattern. If a gene binds a transcription factor which is ubiquitously expressed, then the gene expression will be as well. If the gene binds a transcription factor which is synthesized or active only in a limited number of cells, gene expression will be more cell specific. Regulation of gene expression is for the most part controlled by transcription factors. This regulation provides that the correct gene is activated in the appropriate cell at the precise time for development. Identification of transcription factors and the genes they regulate greatly enhances our understanding of cellular development. As a result intervention methods can be developed to alleviate problems associated with transcription. The present invention addresses this need by providing a novel transcription factor and related compositions and methods.
SUMMARY OF THE INVENTION
The present invention provides a novel testis specific transcription factor and related compositions and methods.
Within one aspect is provided an isolated polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2. Within one embodiment the polypeptide is at least 90% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2. Within another embodiment the polypeptide is covalently linked amino terminally or carboxy terminally to a moiety selected from the group consisting of affinity tags, toxins, radionucleotides, enzymes and fluorophores.
Within another aspect is provided an isolated polynucleotide encoding a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2. Within one embodiment the polypeptide is at least 90% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein said sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2.
Within another aspect the polynucleotide comprising the sequence of nucleotide 1 to nucleotide 1437 of SEQ ID NO:4.
Within yet another aspect is provided an oligonucleotide probe or primer comprising at least 14 contiguous nucleotides of a polynucleotide of SEQ ID NO:4 or a sequence complementary to SEQ ID NO:4.
Also provided by the invention is an expression vector comprising the following operably linked elements: a transcription promoter; a DNA segment encoding a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2; and a transcription terminator. Within one embodiment the DNA segment encodes a polypeptide that is at least 90% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2. Within another embodiment the DNA segment encodes a polypeptide covalently linked amino terminally or carboxy terminally to an affinity tag. Within still another embodiment the DNA segment further encodes a secretory signal sequence operably linked to the polypeptide.
Within another aspect is provided a cultured cell into which has been introduced an expression vector comprising the following operably linked elements: a transcription promoter; a DNA segment encoding a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2; and a transcription terminator; wherein the cell expresses the polypeptide encoded by the DNA segment.
Within another aspect is method of producing a polypeptide comprising: culturing a cell into which has been introduced an expression vector comprising the following operably linked elements: a transcription promoter; a DNA segment encoding a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2; and a transcription terminator; whereby the cell expresses the polypeptide encoded by the DNA segment; and recovering the expressed polypeptide.
Another aspect provided herein is a pharmaceutical composition comprising a polypeptide, the polypeptide comprising a sequence of amino acid residues that is at least 80 identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2; in combination with a pharmaceutically acceptable vehicle.
Within another aspect is an antibody that specifically binds to an epitope of a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2.
Also provided is a binding protein that specifically binds to an epitope of a polypeptide comprising a sequence of amino acid residues that is at least 80% identical in amino acid sequence to residues 1-479 of SEQ ID NO:2, wherein the sequence comprises a POZ domain corresponding to amino acid residues 61-178 of SEQ ID NO:2.
Also provided is a method for detecting a genetic abnormality in a patient, comprising: obtaining a genetic sample from a patient; incubating the genetic sample with a polynucle
Deisher Theresa A.
Yee David P.
Lingenfelter Susan E.
McKelvey Terry
ZymoGenetics Inc.
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