Isolation and Analysis of Human Kekkon-Like Molecules a Family of Potential Inhibitors of ERbB Receptor Tyrosine Kinases
Abstract
Identification and characterization of proteins involved in intrinsic negative feedback loops autoregulating the transforming c-ErbB2/EGF Receptor activity in vivo. Scope: Receptor Tyronsine Kinase (RTK) activation trigger two distinct signal transduction cascades: Mitogenic Ras dependent NAP Kinase activation and P13 Kinase dependent Akt/PKB survival signaling. To date, the regulatory circuits in the Ras signaling pathway are well described. In contrast, no genes involved in the feedback regulating of the PI3K - Akt/PKB signaling branch are known so far. To this end, a cell based genome wide screen employing double stranded RNA interference (dsRNAi) as well as an analysis of Akt mediated transcriptional response using DNA microarrays covering the whole genome have been initiated. Progress: A cuantitative assay to measure amounts of phosphorylated ("active") Akt has been established. The description and analysis of regulatory feedback regulation within the Akt signal transduction pathway using dsRNAi, chemical inhibitors and metabolic perturbation ahs been accomplished. The first genome wide dsRNAi screen for activators of Akt has been completed in replicates. Secondary screens to retest the results form the high-throughput efforts are developed. The data analysis is ongoing.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2004
- Accession Number
- ADA425686
Entities
People
- Lutz R. Kockel
- Norbert Perrimon
Organizations
- Harvard Medical School