Our laboratory has recently shown that induction of expression of the c-fos gene is a cellular response to a wide range of transmembrane signaling agents including growth factors, phorbol esters, and neurotransmitters. We proposed to study properties of the fos protein and its induction. Specifically we propose: 1. To determine the time course of synthesis of fos protein and the association of fos with a second cellular protein, p39 following growth factor induction in 3T3 cells. 2. To determine the molecular weight of the complex of fos protein with p39. From changes in this stoichiometry we will deduce the dynamics of fos-p39 complex formation. 3. To mutagenize the fos protein. We will determine the binding site on fos for p39 and compare this with sites of mutation which alter fos protein transforming ability, nuclear localization, post-translational modification and potential for autoregulation of expression. This will develop a functional topology for the fos protein. 4. To purify the p39 protein through isolation of its complex with fos with the objective of preparing reagents to study p39 expression and structure. 5. To assay the fos protein for a negative regulatory role acting at the fos promoter. 6. To assay other growth factor induced early response genes for fos transcription repressor activity using the antisense technique. 7. To identify proteins interacting with fos regulatory sequences using a novel DNA label transfer assay.
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