Nuclear factors associated with transcriptional regulation

An assay for identifying a compound which modulates the cellular activity of an NF-kB protein, comprising

• (a) contacting a mixture comprising an NF-kB protein and an IkB protein with a test agent;
• and
• (b) detecting the level of NF-kB--IkB protein complexes in the presence of the test agent,
• wherein a difference in the level of NF-kB--IkB protein complexes in the presence of the test agent, relative to the absence of the test agent, indicates that the test agent is a compound which modulates the activity of NF-kB.

An assay for identifying a compound which modulates the NF-kB inhibitory activity of an IkB protein, comprising

• (a) contacting an IkB polypeptide with a test agent under conditions wherein, in the absence of the test agent, the IkB polypeptide is phosphorylated;
• and
• (b) detecting the level of phosphorylation of the IkB polypeptide in the presence of the test agent,
• wherein a difference in the level of phosphorylation of the IkB polypeptide in the presence of the test agent, relative to the absence of the test agent, indicates that the test agent is a compound which modulates the NF-kB inhibitory activity of an IkB protein.

An assay for identifying a compound which modulates the NF-kB inhibitory activity of an IkB protein, comprising

• (a) contacting an IkB polypeptide with a test agent;
• and
• (b) detecting the level of the IkB polypeptide in the presence of the test agent,
• wherein a difference in the level of the IkB polypeptide in the presence of the test agent, relative to the absence of the test agent, indicates that the test agent is a compound which modulates the NF-kB inhibitory activity of an IkB protein.

An assay for identifying a compound which modulates the cellular activity of NF-kB by modulating the translocation of NF-kB to a cell's nucleus, comprising

• (a) contacting a cell expressing an NF-kB polypeptide with a test agent;
• and
• (b) detecting the cellular localization of the NF-kB polypeptide,
• wherein a higher level of NF-kB in the nucleus of the cell in the presence of the test agent, relative to the absence of the test agent, indicates that the test agent is a compound which activates NF-kB, whereas a lower level of NF-kB in the nucleus of the cell in the presence of the test agent, relative to the absence of the test agent, indicates that the test agent is a compound which inhibits NF-kB.

An assay for identifying a compound which modulates the activity of NF-kB, comprising

• (a) contacting a cell having a reporter gene, the transcription of which is dependent upon the activity of NF-kB, with a test agent;
• and
• (b) detecting the level of expression of the reporter gene in the presence of the test agent,
• wherein a difference in the level of expression of the reporter gene in the presence and in the absence of the test agent indicates that the test agent is a compound which modulates the activity of NF-kB.

An assay for identifying a compound which modulates the activity of NF-kB, comprising

• (a) contacting an NF-kB regulatory element with an NF-kB polypeptide;
• and
• (b) detecting the level of interaction between the regulatory element and the NF-kB polypeptide in the presence of the test agent,
• wherein a difference in the level of interaction between the regulatory element and the NF-kB polypeptide in the presence and in the absence of the test agent indicates that the test agent is a compound which modulates the activity of NF-kB.

An assay for identifying a compound which modulates the cellular activity of NF-kB by modulating the level of IkB, comprising

• (a) contacting an IkB polypeptide with a cell extract and a test agent;
• and
• (b) determining the level of IkB polypeptide after contact with the cell extract and the test agent relative to the level of IkB polypeptide after contact with the cell extract alone,
• wherein the presence of a higher level of the IkB polypeptide after contact with the cell extract and the test agent relative to the level of the IkB polypeptide after contact with the cell extract alone, indicates that the test agent is a compound which increases the level of the IkB polypeptide and thereby decreases the activity of NF-kB, whereas the presence of a lower level of IkB polypeptide after contact with the cell extract and the test agent relative to the level of the IkB polypeptide after contact with the cell extract alone, indicates that the test agent is a compound which reduces the level of IkB and thereby increases the activity of NF-kB.

An assay for identifying a compound which modulates the cellular activity of NF-kB by modulating the phosphorylation of IkB, comprising

• (a) contacting a cell or cell lysate comprising an IkB polypeptide with a test agent;
• and
• (b) determining the level of phosphorylation of the IkB polypeptide in the cell or cell lysate contacted with the test agent relative to a cell or cell lysate which was not contacted with the test agent,
• wherein the presence of a higher level of phosphorylated IkB polypeptide in the cell or cell lysate contacted with the test agent relative to a cell or cell lysate which was not contacted with the test agent, indicates that the test agent is a compound which increases the phosphorylation of the IkB polypeptide and thereby increases the activity of NF-kB, whereas the presence of a lower level of phosphorylated IkB polypeptide in the cell or cell lysate contacted with the test agent, relative to the cell or cell lysate which was not contacted with the test agent, indicates that the test agent is a compound which reduces the phosphorylation of IkB and thereby decreases the activity of NF-kB.

An assay for identifying a compound which modulates the cellular activity of NF-kB by modulating the phosphorylation of IkB, comprising

• (a) contacting an IkB polypeptide with a cell extract and a test agent;
• and
• (b) determining the level of IkB phosphorylation after contact with the cell extract and the test agent relative to the level of IkB phosphorylation after contact with the cell extract alone,
• wherein the presence of a higher level of IkB phosphorylation after contact with the cell extract and the test agent relative to the level of IkB phosphorylation after contact with the cell extract alone, indicates that the test agent is a compound which increases the phosphorylation of the IkB polypeptide and thereby increases the activity of NF-kB, whereas the presence of a lower level of IkB phosphorylation after contact with the cell extract and the test agent relative to the level of IkB phosphorylation after contact with the cell extract alone, indicates that the test agent is a compound which reduces the phosphorylation of IkB and thereby decreases the activity of NF-kB.

An assay for identifying a compound which modulates the cellular activity of NF-kB by modulating the level of expression of an IkB gene, comprising

• (a) contacting a cell or cell lysate comprising an IkB gene with a test agent;
• and
• (b) determining the level of expression of the IkB gene in the cell or cell lysate contacted with the test agent relative to a cell or cell lysate which was not contacted with the test agent,
• wherein the presence of a higher level of expression of an IkB gene in the cell or cell lysate contacted with the test agent relative to a cell or cell lysate which was not contacted with the test agent, indicates that the test agent is a compound which increases the expression of the IkB gene and thereby decreases the activity of NF-kB, whereas the presence of a lower level of expression of the IkB gene in the cell or cell lysate contacted with the test agent, relative to the cell or cell lysate which was not contacted with the test agent, indicates that the test agent is a compound which reduces the level of expression of the IkB gene and thereby increases the activity of NF-kB.