The protein-tyrosine kinase Fer and the highly homologous proto-oncoprotein Fps/Fes are implicated in signaling from a variety of growth factor and cytokine receptors. Here we examine the molecular basis of Fer kinase activation with an emphasis on the role of oligomerization. We show that Fer forms trimers in vivo and that disruption of either the first or second coiled-coil domain abolishes oligomerization, suggesting a cooperative interaction between these two domains. Although Fps/Fes also forms homotypic oligomers, probably via homologous coiled-coil domains, no heterotypic interactions were observed between Fer and Fps/Fes. Incorporation of catalytically inactive Fer peptides into the oligomeric complex caused only mild reduction of wild type Fer kinase activity, suggesting that kinase-inactive Fer would not behave as a potent dominant negative. Although oligomerization of Fer can potentiate autophosphorylation in trans at three major phosphorylation sites, these residues can likely also be phosphorylated in cis. In contrast, the testis-specific FerT isomer does not oligomerize and is able to autophosphorylate in cis at two of the same three residues autophosphorylated in Fer. These results suggest that although oligomerization potentiates autophosphorylation in trans, this is apparently not necessary for Fer activation.