The Salk Institute for Biological Studies

Research Development Foundation

The Salk Institute

Strang Cancer Prevention Center

Marc R Montminy

James N Leonard

Seema Sharma

In accordance with the present invention, it has been discovered that CREB binding protein (CBP) cooperates with upstream activators involved in the activation of transcription of such signal dependent transcription factors as c-Jun (responsive to phorbol ester), serum response factor, and the like. It has also been discovered that CBP can be employed in an assay to identify compounds which disrupt the ability of such signal dependent transcription factors to activate transcription. In another aspect, it has been discovered that CBP can be employed in an assay to identify new signal dependent transcription factors. In yet another aspect of the present invention, it has been discovered that CBP can be employed in an assay to identify novel co-factor protein(s) which mediate the interaction between signal dependent transcription factors and inducer molecules involved in the activation of transcription. Accordingly, the present invention provides methods for the identification of compounds which inhibit activation of cAMP and mitogen responsive genes and methods for the identification of novel signal dependent transcription factors and co-factor proteins. In still another aspect, methods employing compounds which inhibit intracellular signal-induced response pathways have been developed for the treatment of diabetes mellitus.

Identification of transcription factor within a cAMP-responsive transcriptional enhancer binding protein (CREB), and uses therefore

Cyclic amp response element binding protein (CREB)-CREB binding protein

One embodiment of the present invention provides a method of testing for compounds which may induce STF-1 transcription. Pancreatic islet cells which express the STF-1/lacz fusion gene are isolated and the effect of various compounds on STF-1 expression is measured by adding the compound of interest to STF-1/lacZ expressing cells. LacZ activity in control and treated cells is then quantitated by a colorimetric assay. Using this method, a large number of compounds can be screened and STF-1 inducing compounds can be identified readily. Another embodiment of the present invention provides a method of using the STF-1 promoter to mark insulin producing pancreatic islet cells in vivo. In this regard, the green fluorescent protein (GFP) served as an indicator. The introduction of the STF-1 green fluorescent protein transgene into pigs allows for the efficient and rapid recovery of insulin producing cells from the pancreas.

Methods for the characterization of compounds which stimulate STF-1

The invention relates to a homeoprotein regulator of insulin gene expression having the characteristics of: binding to an element of an insulin gene promoter; being modulated by a Ca.sup.++ -dependent CaM kinase IV; and having homology to a nucleotide sequence encoded by a Hox gene complex. Also included within the invention are DNA sequences encoding the homeoprotein regulators of insulin gene expression, antibodies directed to the homeoprotein regulators of insulin gene expression, and diagnostic and therapeutic materials and utilities for the homeoprotein regulators of insulin gene expression.

Islet-specific homeoprotein and transcriptional regulator of insulin

Phospho-specific antibodies which recognize the phosphorylated forms of cAMP-responsive transcription factors are provided.

Phosphospecific transcription factor antibodies

In accordance with the present invention, it has been discovered that CREB binding protein (CBP) cooperates with upstream activators involved in the activation of transcription of such signal dependent transcription factors as c-Jun (responsive to phorbol ester), serum response factor, and the like. It has also been discovered that CBP can be employed in an assay to identify compounds which disrupt the ability of such signal dependent transcription factors to activate transcription. In another aspect, it has been discovered that CBP can be employed in an assay to identify new signal dependent transcription factors. In yet another aspect of the present invention, it has been discovered that CBP can be employed in an assay to identify novel co-factor protein(s) which mediate the interaction between signal dependent transcription factors and inducer molecules involved in the activation of transcription. Accordingly, the present invention provides methods for the identification of compounds which inhibit activation of cAMP and mitogen responsive genes and methods for the identification of novel signal dependent transcription factors and co-factor proteins.

Assays for the identification of compounds which inhibit activation of

In accordance with the present invention, there are provided novel homeobox-type pancreatic islet transcription factor proteins useful to bind to tissue-specific elements (TSEs) within a pancreatic islet hormone gene promoter and modulate hormone gene expression both in vivo and in vitro. Nucleic acid sequences encoding such transcription factor proteins and assays employing same are also disclosed. The invention transcription factor proteins can be employed in a variety of ways, for example, to modulate RNA transcription, for production of antibodies thereto, in therapeutic compositions and methods employing such proteins and/or antibodies.

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Encinitas, CA, United States of America

Marc R Montminy