The neuropeptide bombesin is known for its potent mitogenic activity on murine 3T3 fibroblasts and other cells. Recently it has been implicated in the pathogenesis of small cell lung carcinoma, in which it acts through an autocrine loop of growth stimulation. Phosphotyrosine (P-Tyr) antibodies have been Many recent reports on proliferative growth factors provide evidence that peptides already known for their hormonal or neurotransmitter nature can act as mitogens. Two mammalian members of the tachykinin family, substance P and substance K, which are present in sensory neurons, specifically stimulate DNA synthesis in cultured arterial smooth muscle cells and human skin fibroblasts (32). The neurohormone vasopressin and the tetradecapeptide bombesin (1) are also potent mitogens. The former stimulates DNA synthesis in chondrocytes (26), bone marrow cells (18), and Swiss 3T3 mouse fibroblasts (34). The latter, which stimulates hormone secretion (37) and smooth muscle contraction and affects temperature control and behavior in mammals (14, 38), shows potent mitogenic activity on Swiss 3T3 fibroblasts (35), in which high-affinity receptors for peptides of the bombesin family were recently demonstrated (45).Bombesin and bombesinlike peptides have been detected in various normal mammalian tissues (28,40,42), together with specific cell surface receptors (21, 27). These peptides have also been shown to behave as growth factors for normal human bronchial epithelial cells (44) and for human small cell lung carcinoma, in which they appear to act in an autocrine circuit stimulating growth (6). Furthermore, high levels of bombesinlike immunoreactivity were reported in rat mammary (13) human lung (10, 29), and human thyroid (25) tumors. The broad tissue specificity of this peptide and its mitogenic activity on different cell types suggest a role for bombesin in growth control of both normal and transformed cells. oncogenes (16,17). Bombesin, like the tachykinins and vasopressin, stimulates polyphosphoinositide breakdown (2). It has also recently been shown that bombesin induces rapid and transient expression of c-fos and c-myc protooncogenes (A. P. Palumbo, P. Rossino, and P. M. Comoglio, Exp. Cell Res., in press).To assess whether the mitogenic effect of bombesin involves a receptor-linked protein tyrosine kinase activity, we used purified antibodies capable of recognizing phosphotyrosine (P-Tyr) residues in proteins (5, 31, 46) (P-Tyr antibodies). These antibodies have been used to detect tyrosine protein kinases associated with growth factor receptors through identification of the autophosphorylated receptor and its cellular substrates in mitogen-stimulated cells (7a, 9, 11, 12, 43). The present investigation demonstrates that bombesin specifically stimulates the phosphorylation on tyrosine of a membrane protein of Mr 115,000 (p115). Immunoprecipitates of this protein, made with P-Tyr antibodies from bombesin-stimulated cells, displayed a bombesindependent tyrosine kinase activity and bound labeled bombesin in a specific...