Molecular Cloning Of Human Paxillin, a Focal Adhesion Protein Phosphorylated by P210
(*)
- Ravi Salgia(1),
- Jian-Liang Li(2),
- Su Hao Lo(2),
- Beatrice Brunkhorst(2),
- Geoffrey S. Kansas(1),
- E. Sholeh Sobhany(3),
- Yaping Sun(2),
- Evan Pisick(1),
- Michael Hallek(1),
- Timothy Ernst(1),
- Ramana Tantravahi(3),
- Lan Bo Chen(2) and
- James D. Griffin(1)(§)
- From the (1)Division of Hematologic Malignancies, the
- (2)Division of Cellular and Molecular Biology, and the
- (3)Division of Cytogenetics, Dana-Farber Cancer Institute, Boston, Massachusetts 02115
- §To whom correspondence should be addressed: Division of Hematologic Malignancies, Dana-Farber Cancer Institute, 44 Binney St., Boston, MA 02115. Tel.: 617-632-3360; Fax: 617-632-4388.
Abstract
Paxillin is a 68-kDa focal adhesion protein that is phosphorylated on tyrosine residues in fibroblasts in response to transformation
by v-src, treatment with platelet-derived growth factor, or cross-linking of integrins. Paxillin has been shown to have binding sites
for the SH3 domain of Src and the SH2 domain of Crk in vitro and to coprecipitate with two other focal adhesion proteins, vinculin and focal adhesion kinase (p125
). After preliminary studies showed that paxillin was a substrate for the hematopoietic oncogene p210
, we investigated the role of this protein in hematopoietic cell transformation and signal transduction. A full-length cDNA
encoding human paxillin was cloned, revealing multiple protein domains, including four tandem LIM domains, a proline-rich
domain containing a consensus SH3 binding site, and three potential Crk-SH2 binding sites. The paxillin gene was localized
to chromosome 12q24 by fluorescence in situ hybridization analysis. A chicken paxillin cDNA was also cloned and is predicted to encode a protein approximately 90% identical
to human paxil-lin. Paxillin coprecipitated with p210
and mul-tiple other cellular proteins in myeloid cell lines, suggesting the formation of multimeric complexes. In normal
hematopoietic cells and myeloid cell lines, tyrosine phosphorylation of paxillin and coprecipitation with other cellular proteins
was rapidly and transiently induced by interleukin-3 and several other hematopoietic growth factors. The predicted structure
of paxillin implicates this molecule in protein-protein interactions involved in signal transduction from growth factor receptors
and the BCR/ABL oncogene fusion protein to the cytoskeleton.
Footnotes
-
↵* This work was supported in part by National Institutes of Health Grants CA 60821, CA36167, and GM 3818. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore by 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) U14588 [GenBank]and U14589[GenBank].
-
↵1 The abbreviations used are:
- IL
-
interleukin
- kb
-
kilobase(s)
- PCR
-
polymerase chain reaction
- FISH
-
fluorescence in situ hybridization
- GM-CSF
-
granulocyte-macrophage-colony stimulating factor
- RACE
-
rapid amplification of cDNA ends
- GST
-
glutathione S-transferase
- PAGE
-
polyacrylamide gel electrophoresis
- CML
-
chronic myelogenous leukemia.
-
- Received October 31, 1994.
- Revision received December 12, 1994.
- © 1995 by The American Society for Biochemistry and Molecular Biology, Inc.
