Characterization of Schwannomin Role in Protein Translation
Abstract
We demonstrated that schwannomin interacts with the eukaryotic initiation factor 3 (eIF3) p110 subunit eight and now investigate the role of schwannomin in eIF3 function. eIF3 is required for the 40 S ribosomal subunit bound to the ternary complex (eIF2-GTP-methionine) to interact with the 5' end of the mRNA, and is required for progression of protein translation. Keeping with our Objectives and Statement of Work, we have: (1) verified the interaction of schwannomin and p110 by using the non-transcriptionally-based ras-rescue yeast two-hybrid system, (2) produced and validated two high-quality p110 antibodies, (3) determined the cross-reactivity of our anti-p110 antibodies in mouse rat and human cells, (4) co-immunoprecipitated schwannomin and p110 using one of the new p110 antibodies, (5) cloned the full-length p110 cDNA and validated its expression, (6) co-localized p110 and schwannomin in STS26T schwannoma cells by confocal microscopy, (7) developed a NF2 inducible tet-off MEF cell line for the study on p110 function, (8) narrowed the region of p110 interaction in schwannomin to p110 amino acids 327-635 by yeast two-hybrid, (9) cloned eight plasmids for further narrowing of the p110 region of interaction with schwannomin by in vitro methods, and (10) showed the effects of NF2 mutations_on p110 interaction by the yeast two-hybrid method.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2001
- Accession Number
- ADA400194
Entities
People
- Daniel R. Scoles
Organizations
- Cedars-Sinai Medical Center