By yeast two-hybrid screening with the Src kinase Lyn as bait, we identified a novel gene product with features of a scaffolding protein. Reported as Felic (Fes-related, with homology to Ezrin, Lyn interactor with Cdc42), it is related to the CIP4 (Cdc42 Interacting Protein-4) gene. Southern blotting for CIP4/Felic of genomic DNA shows a single band, suggesting no gene duplication. Felic differs from CIP4 because of a 29 nucleotide sequence derived from the 3' end of intron 13. Consequently, there is an out-of-frame translation that destroys an SH3 domain. Analysis of various tissues shows that the original CIP4 is the predominant transcript. Therefore, we propose to call that, CIP4a and Felic, CIP4b. During screening of the colorectal CaCo2 cell line, clones corresponding to a third CIP4-related transcript (CIP4c) were identified. CIP4c encodes a premature stop codon, resulting in the loss of the SH3 domain. A fourth, relatively abundant transcript (CIP4h) was isolated from heart, lung, and trachea tissue. CIP4h retains the SH3 domain. CIP4 levels are modified by all-trans-retinoic acid. The presence of alternative splice transcripts, with or without SH3 domains, suggests that CIP4 regulates cytoskeletal organization through structural-functional differences in a tissue-specific manner. (C) 2002 Elsevier Science (USA). All rights reserved.