Novel receptors: their identification, characterization, preparation and use

Claim 2

An assay for screening and identifying materials having a putative potential of binding to a hormone or hormone-like receptor comprising the steps of: (a) providing a hormone or hormone-­like receptor species in a form having improved trans-­activation transcription activity, (b) challenging the form of receptor species with one or more of a battery of test materials having putative potential of binding to a hormone or hormone-like receptor, (c) monitoring the effect of the test material by measuring the amount of transcription induced by the receptor species, and (d) selecting candidates from the battery of test materials capable of having an effect on the trans-activation transcription activity of said receptor species.

Claim 3

An assay as claimed in Claim 1 further comprising the additional step following step (d), comprising: (e) employing the candidate in the preparation of a composition containing the candidate as an essential component, the composition being useful to impart its biofunction properties on a corresponding receptor when it is contacted in vivo with the receptor.

Claim 4

An assay as claimed in Claim 2 further comprising the additional step following step (e), comprising: (f) contacting the composition with a human subject.

Claim 5

A hormone or hormone-like receptor as a polypeptide having increased trans-activation transcription activity of a promoter with which it is associated, by virtue of its intrinsic ability to bind to a DNA sequence response element of the promoter or by its ability to associate with other polypeptide that bind to the DNA sequence response element, wherein the receptor contains a plurality of trans-activation transcription domains thereof, of a functional fragment thereof, located within the molecule in a location outside of its DNA-binding and ligand-binding domains.

Claim 6

A hormone or hormone-like receptor as a polypeptide having increased trans-activation transcription activity of a promoter with which it is associated, by virtue of its intrinsic ability to bind to a DNA sequence response element of the promoter or by its ability to associate with other polypeptide that bind to the DNA sequence response element, wherein the receptor contains a plurality of trans-activation transcription domains thereof, of a functional fragment thereof, located within the molecule in a location outside of its DNA-binding and ligand-binding domains.

Claim 7

A receptor as claimed in Claim 4 wherein the receptor is based on the human glucocorticoid receptor.

Claim 8

A receptor as claimed in Claim 4 wherein the trans-activation transcription domain is the τ₁ domain.

Claim 9

A receptor as claimed in Claim 6 wherein the receptor contains two of the τ₁ domains.

Claim 10

A receptor as claimed in Claim 7 wherein the second of the τ₁ domains is located adjacent to the first.

Claim 11

A receptor as claimed in Claim 7 wherein the second of the τ₁ domains is located in a C-terminal distal location from the first τ₁ sequence.

Claim 12

A receptor as claimed in any one of Claims 4 to 9 lacking a ligand-binding domain.

Claim 13

A species of a receptor as claimed in Claim 4 which is human G442 glucocorticoid receptor.

Claim 14

A hormone or hormone-like receptor having trans-activation transcription domains additional to that of the parent receptor, autonomous with both the DNA-binding and ligand-binding domains of the parent receptor, the additional trans-activation transcription domains being located in a position outside of each of the DNA-binding and ligand-binding domains, if present, and having a DNA-­binding biofunctionality.

Claim 15

A hormone or hormone-like receptor having trans-activation transcription domains additional to that of the parent receptor, autonomous with both the DNA-binding and ligand-binding domains of the parent receptor, the additional trans-activation transcription domains being located in a position outside of each of the DNA-binding and ligand-binding domains, if present, and having a DNA-­binding biofunctionality.

Claim 16

A DNA molecule that is a recombinant DNA molecule or a cDNA molecule encoding a receptor as claimed in Claim 12.

Claim 17

An expression vector operatively harboring DNA encoding a receptor as claimed in Claim 13.

Claim 18

A recombinant host cell transfected with an expression vector as claimed in Claim 14.

Claim 19

A cell culture comprising cells as claimed in claim 15 and an extrinsic support medium assuring the viability of the cells.

Claim 20

A process of preparing a human receptor as claimed-in Claim 12 which comprises expressing in a recombinant host cell transfecting DNA encoding the receptor.

Claim 21

A process which comprises recovering and purifying a receptor as claimed in Claim 12 to a form having quality and quantity sufficient to enable its use in assays that enable measurement of extrinsically induced biofunctionality of the receptor.

Claim 22

In a bioassay for determining the functionality of a hormone or hormone-like receptor, the bioassay comprising: (a) transfecting into receptor negative cells one or more expression vectors harboring an operative hormone responsive promoter/enhancer element functionally linked to an operative reporter or other desired DNA and an operative DNA sequence encoding the receptor, (b) culturing the transfected cells from step (a) in the presence or absence of extrinsic material having the putative ability to activate the hormone responsive promoter/enhancer element, (c) monitoring in the cells induction of the product of the reporter or desired DNA sequence, and (d) measuring in the cells the expression of reporter or other desired DNA, the improvement wherein the hormone or hormone-like receptor is one having increased trans-activation transcription activity compared with the parent receptor, the activity additional to the parent receptor by virtue of domains, or functional fragments thereof, located within the molecule at a position outside of each of a DNA-­binding and ligand-binding domain of said receptor.

Claim 23

In a bioassay for determining the functionality of a hormone or hormone-like receptor, the bioassay comprising: (a) transfecting into receptor negative cells one or more expression vectors harboring an operative hormone responsive promoter/enhancer element functionally linked to an operative reporter or other desired DNA and an operative DNA sequence encoding the receptor, (b) culturing the transfected cells from step (a) in the presence or absence of extrinsic material having the putative ability to activate the hormone responsive promoter/enhancer element, (c) monitoring in the cells induction of the product of the reporter or desired DNA sequence, and (d) measuring in the cells the expression of reporter or other desired DNA, the improvement wherein the hormone or hormone-like receptor is one having increased trans-activation transcription activity compared with the parent receptor, the activity additional to the parent receptor by virtue of domains, or functional fragments thereof, located within the molecule at a position outside of each of a DNA-­binding and ligand-binding domain of said receptor.