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Molecular and Cellular Biology, October 1998, p. 5838-5851, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification of a Novel Cortactin SH3 Domain-Binding Protein and Its Localization to Growth Cones of Cultured Neurons

Yunrui Du, Scott A. Weed, Wen-Cheng Xiong, Trudy D. Marshall,dagger and J. Thomas Parsons*

Department of Microbiology and Cancer Center, University of Virginia Health Science Center, Charlottesville, Virginia 22908

Received 23 April 1998/Returned for modification 21 May 1998/Accepted 18 June 1998

Cortactin is an actin-binding protein that contains several potential signaling motifs including a Src homology 3 (SH3) domain at the distal C terminus. Translocation of cortactin to specific cortical actin structures and hyperphosphorylation of cortactin on tyrosine have been associated with the cortical cytoskeleton reorganization induced by a variety of cellular stimuli. The function of cortactin in these processes is largely unknown in part due to the lack of information about cellular binding partners for cortactin. Here we report the identification of a novel cortactin-binding protein of approximately 180 kDa by yeast two-hybrid interaction screening. The interaction of cortactin with this 180-kDa protein was confirmed by both in vitro and in vivo methods, and the SH3 domain of cortactin was found to direct this interaction. Since this protein represents the first reported natural ligand for the cortactin SH3 domain, we designated it CortBP1 for cortactin-binding protein 1. CortBP1 contains two recognizable sequence motifs within its C-terminal region, including a consensus sequence for cortactin SH3 domain-binding peptides and a sterile alpha motif. Northern and Western blot analysis indicated that CortBP1 is expressed predominately in brain tissue. Immunofluorescence studies revealed colocalization of CortBP1 with cortactin and cortical actin filaments in lamellipodia and membrane ruffles in fibroblasts expressing CortBP1. Colocalization of endogenous CortBP1 and cortactin was also observed in growth cones of developing hippocampal neurons, implicating CortBP1 and cortactin in cytoskeleton reorganization during neurite outgrowth.

* Corresponding author. Mailing address: Department of Microbiology, Box 441, Health Science Center, University of Virginia, Charlottesville, VA 22908. Phone: (804) 924-5395. Fax: (804) 982-1071. E-mail: jtp{at}virginia.edu.

dagger Present address: c/o CMHA (NF Division), St. John's, Newfoundland A1C 5X3, Canada.

Molecular and Cellular Biology, October 1998, p. 5838-5851, Vol. 18, No. 10
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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