Identification of novel factors that block programmed cell death or apoptosis by targeting jnk

Abstract

Methods and compositions for modulating apoptosis by acting on the c-Jun-N-terminal kinase (JNK) pathway and assays for the isolation of agents capable of modulating apoptosis, including modulators of the JNK pathway are disclosed. A method of modulating JNK pathway independent of Gadd46β is disclosed. Methods and compositions are presented for the preparation and use of novel therapeutic compositions for modulating diseases and conditions associated with elevated or decreased apoptosis.

1 . A method for screening and identifying an agent that modulates activity of the JNK pathway in vivo, the method comprising:
 (a) obtaining a candidate agent that binds to a factor which binds to a molecule with an amino acid sequence consisting essentially of NH2-TGHVIAVKQMRRSGNKEENKRILMD-COOH (SEQ ID NO: 1);   (b) administering the agent to an animal; and   (a) determining whether the level of JNK activity or programmed cell death in the animal is increased compared to JNK activity or programmed cell death in animals not receiving the agent.   
     
     
         2 . A method to identify inhibitors of Gadd45β, the method comprising:
 (a) screening for a candidate compound that binds to peptidic regions consisting essentially of amino acid sequences from positions 60-86 (AIDEEEEDDIALQIHFTLIQSFCCDND, SEQ ID NO: 2) and 69-86 (IALQIHFTLIQSFCCDND, SEQ ID NO: 3) of Gadd45β; and   (b) determining the ability of the candidate compound to bind to Gadd45β or interfere with Gadd45β-mediated inhibition of JNKK2.   
     
     
         3 . A method to identify agents that interfere with binding of JNKK2 to a molecule capable of binding to positions 142-166 (TGHVIAVKQMRRSGNKEENKRILMD, SEQ ID NO: 1) of the full length JNKK2, the method comprising:
 (a) obtaining an agent that interferes with the binding of the molecule to positions 142-166 (TGHVIAVKQMRRSGNKEENKRILMD) of the full length JNKK2;   (b) contacting a cell with the agent under conditions that would induce JNK activation or programmed cell death; and   (c) comparing cells contacted with the agent to cells not contacted with the agent to determine if the JNK pathway is upregulated.