Multiple forms of p55PIK, a regulatory subunit of phosphoinositide 3-kinase, are generated by alternative initiation of translation

A cDNA encoding p55PIK, one of the regulatory subunits of phosphoinositide (phosphatidylinositol) 3-kinase, was cloned from a cDNA library derived fro m the mouse mammary epithelial cell line C57MG. The cDNA coding for full-le ngth p55PIK was transiently expressed in COS-7 cells. Western blot analysis of p55PIK expression using a specific antibody against p55PIK revealed tha t multiple protein products with different molecular masses were detected i n COS-7 cell extracts. Experiments presented here demonstrate that multiple forms of p55PIK detected in COS-7 cells were produced by alternative initi ation of translation. We also show that at least two in-frame start codons (AUG#2 and AUG#5) in p55PIK mRNA are used in COS-7 cells for the initiation of translation of p55PIK into proteins of 54 kDa and 50 kDa respectively, p55PIK mRNA was also alternatively translated into two proteins in PC cells , a mouse teratoma cell line, indicating that the alternative initiation of translation of p55PIK is not restricted to COS-7 cells. Results from immun oprecipitation and Western blot analysis showed that two forms (54 kDa and 50 kDa protein species) of p55PIK were detected in C57MG cells. Interesting ly, when C57MG cells were treated with insulin, only p55PIK, but not p50PIK , bound to insulin receptor substrate-1 protein, providing evidence that di fferent forms of p55PIKs may have specific distinct roles in signal transdu ction pathways.