Abl Family Kinases and Cbl Cooperate with the Nck Adaptor to Modulate Xenopus Development

doi:10.1074/jbc.M005424200

Abl Family Kinases and Cbl Cooperate with the Nck Adaptor to Modulate Xenopus Development*

From the Laboratory of Molecular Medicine, Children's Hospital and Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

Abstract

We previously showed that overexpression of the Nck Src homology (SH) 2/SH3 adaptor in Xenopus embryos induced developmental defects including anterior truncation and mesoderm ventralization. Mutagenic analysis indicated that this was due to relocalization of endogenous proteins that bind the first two SH3 domains of Nck. We therefore screened a Xenopus expression library with Nck SH3 domains to identify Nck-interacting proteins, and evaluated candidate binding proteins for a potential role in Nck-induced anterior truncation/ventralization. Of 39 binding proteins analyzed, only the Abl-related kinase Arg and the Cbl proto-oncogene product bound preferentially to the first two SH3 domains in tandem compared with the individual domains, consistent with a role in the developmental phenotype. High level overexpression of c-Abl or Arg alone induced anterior truncation, as did lower levels of an activated form of Abl; Cbl alone had no effect. In a sensitized system where subthreshold amounts of a ventralizing Nck mutant were expressed, co-expression of the combination of Abl or Arg and Cbl at modest levels strongly potentiated anterior truncation, while Arg, Abl, or Cbl alone were without effect. These results suggest a role for both Cbl and Abl family kinases in patterning the Xenopus embryo.

Footnotes

↵* This work was supported in part by National Institutes of Health Grant CA82258 (to B. J. M.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) and .
↵‡ Current address: Dept. of Anatomy, University of California, San Francisco, CA 94143.
↵§ These two authors contributed equally to this work.
↵¶ Supported by the Howard Hughes Medical Institute.
↵‖ Current address: Dept. of Microbiology and Immunology, Tokyo Women's Medical University, Tokyo 162-8666, Japan.
↵** Current address: Dept. of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030.
↵‡ Current address: First Dept. of Pathology, Hamamatsu University School of Medicine, Hamamatsu 430, Japan.
↵§§ To whom correspondence should be addressed: Dept. of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Ave., Farmington CT 06030. Tel.: 860-679-1836; Fax: 860-679-8345; E-mail: bmayer@neuron.uchc.edu.
Published, JBC Papers in Press, August 30, 2000, DOI 10.1074/jbc.M005424200
↵2 T. Miyoshi-Akiyama, L. M. Aleman, J. M. Smith, C. E. Adler, and B. J. Mayer, submitted for publication.
Abbreviations:

SH

Src homology

PCR

polymerase chain reaction

GST

glutathioneS-transferase

aa

amino acid(s)

DAI

dorsoanterior index

PDGF

platelet-derived growth factor

PAGE

polyacrylamide gel electrophoresis

wt

wild type

HA

hemagglutinin

FGF

fibroblast growth factor
- Received June 21, 2000.
- Revision received August 1, 2000.
The American Society for Biochemistry and Molecular Biology, Inc.