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Nucleic protein 'Shoca'-a component of wnt signal transmission channel

A technology of signal transduction and pathway, applied in the field of signal protein

Inactive Publication Date: 2005-03-23
THE UNIV CHILDRENS HOSPITAL OF BOTH CANTONS OF BASEL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There have been many reports of transcriptional overexpression and sometimes underexpression of wnt genes in human cancers, but mRNA expression levels are only relative

Method used

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  • Nucleic protein 'Shoca'-a component of wnt signal transmission channel
  • Nucleic protein 'Shoca'-a component of wnt signal transmission channel
  • Nucleic protein 'Shoca'-a component of wnt signal transmission channel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0181] Example 1. Shoca-1, a novel gene involved in wnt-mediated signaling.

[0182] The present inventors have recently identified a novel gene in mouse and human tissues, the product of which is directly involved in wnt-mediated signaling. The gene (SEQ ID NO: 1 and SEQ ID NO: 3) encodes the SH2 domain of the adapter protein containing 52kDa (proved by Western blot hybridization experiment), thus the gene is named Shoca (SH2-domain containing adapter protein)- 1. Subsequently, another gene similar to Shoca (SEQ ID NO: 5) was identified by using EST analysis in the common domain. The presence or function of these genes in any species has not been reported so far.

[0183] Mouse Shoca-2 was discovered by a blast search in the EST database using the cDNA sequence of mouse Shoca-1. To obtain the full-length coding sequence, 5' and 3' RACE (rapid amplification of cDNA ends) was performed using partial EST sequences. Afterwards, the full-length cDNA having the same sequence as...

Embodiment 2

[0187] Example 2. Production of Shoca-specific antibodies

[0188] Antibody preparation A

[0189] The anti-Shoca-1 antibody (rabbit polyclonal serum) used to determine the subcellular localization was produced in the inventor's institute. Different batches were used whereby animals were bled at different times after repeated booster immunizations. The immunogen is a mouse Shoca-1 derived peptide coupled to the carrier protein KLH:

[0190] Peptide Shoca #0: NH 2 -CLPDTSPPSPLTGPDRTWERPLRC-CONH 2

[0191] This peptide represents a stretch of 22 amino acids corresponding to residues 272 to 293 of mouse Shoca-1. For booster immunization, N-terminal and C-terminal cysteine ​​residues were introduced to form loops on the protein carrier.

[0192] Antibody preparation B

[0193] The anti-Shoca-1 antibody used in the staining experiment of Example 5 was raised in two rabbits by Eurogentec and is also representative of a polyclonal preparation. All preparations used for staini...

Embodiment 3

[0217] Example 3. Shoca-1 and Shoca-2 expression profiles

[0218] Shoca-1 is expressed in mouse thymocytes as early as when thymic primordia are first detected, and it is also present in the thymus of nude mice. Thus, Shoca-1 expression is independent of thymic lymphopoiesis, and specifically the structural cross-talk between thymocytes and thymic epithelial cells. Shoca-1 expression was also detected in the bone marrow, particularly in mesenchymal cells known to support blood cell production. Although Northern blot analysis (Northern blot analysis) was not sensitive enough to detect specific transcripts, PCR analysis revealed that brain, kidney and lung were other sites where Shoca-1 was expressed in low abundance (Fig. 6). In contrast, Shoca-2 expression was more common in different tissues (Fig. 6).

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Abstract

An isolated Shoca polypeptide comprising: (i) the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4; or (ii) a variant thereof which is capable of interacting with a polypeptide of the wnt signalling pathway; or (iii) a fragment of (i) or (ii) which is capable of interacting with a polypeptide of the wnt signalling pathway, a polypeptide encoding said polypeptide, a method for identifying a modulator of the wnt signalling pathway utilising said polypeptide or polynucleotide and methods of diagnosing and treating cancer.

Description

technical field [0001] The present invention relates to a novel signaling protein associated with healthy, neoplastic and tumorigenic tissues. The present invention also relates to screening methods for identifying modulators of the activity and / or expression of the signaling protein, and to methods related to the diagnosis, prognosis and treatment of cancer. Background technique [0002] Cancer in humans arises from the accumulation of independent genetic variations that affect the transcriptional programs normally responsible for controlling cell growth and survival. Within a single clinical category, cancers can appear to exhibit disparate genetic defects that are, however, part of a shared signal transduction pathway. The discovery of the Wnt signaling pathway (summarized in Figure 1) and the structure / function analysis of its unique molecular components provides a striking example for identifying shared features in tumorigenesis (Polakis (2000) Genes and Genes & Devel...

Claims

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Application Information

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IPC IPC(8): C12N15/09A61K38/00A61K38/16A61K45/00A61K47/48A61K51/10A61P35/00C07K14/47C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/12C12N15/63C12Q1/02C12Q1/68G01N33/53
CPCC07K14/47A61K51/1093A61K38/00C07K16/18A61K47/48707A61K47/6887A61P35/00
Inventor 乔治·安德里亚·荷兰德尔
Owner THE UNIV CHILDRENS HOSPITAL OF BOTH CANTONS OF BASEL
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