The Rho-specific Guanine Nucleotide Exchange Factor Dbs Regulates Breast Cancer Cell Migration

• Published in: The Journal of biological chemistry Vol. 284; no. 23; pp. 15771 - 15780
• Main Authors: Liu, Zhuoming, Adams, Homer C., Whitehead, Ian P.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.06.2009; American Society for Biochemistry and Molecular Biology
• Subjects: Breast Neoplasms - genetics; Breast Neoplasms - physiopathology; cdc42 GTP-Binding Protein - genetics; Cell Line, Tumor; Cell Movement; Crk-Associated Substrate Protein - genetics; Female; Gene Expression Regulation, Neoplastic; Guanine Nucleotide Exchange Factors - genetics; Guanine Nucleotide Exchange Factors - physiology; Humans; Mechanisms of Signal Transduction; Neoplasm Proteins - genetics; Neoplasm Proteins - physiology; Protein-Tyrosine Kinases - genetics; Rho Guanine Nucleotide Exchange Factors; Schwann Cells - physiology; Suppression, Genetic
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M901853200

Dbs is a Rho-specific guanine nucleotide exchange factor (RhoGEF) that regulates neurotrophin-3-induced cell migration in Schwann cells. Here we report that Dbs regulates cell motility in tumor-derived, human breast epithelial cells through activation of Cdc42 and Rac1. Cdc42 and Rac1 are activated in T47D cells that stably express onco- or proto-Dbs, and activation is dependent upon growth of the cells on collagen I. Transient suppression of expression of Cdc42 or Rac1 by small interfering RNAs attenuates Dbs-enhanced motility. Both onco- and proto-Dbs-enhanced motility correlates with an increase in tyrosine phosphorylation of focal adhesion kinase on Tyr-397 and p130Cas on Tyr-410 and an increase in the abundance of the Crk·p130Cas complex. Suppression of expression of Cdc42 or its effector, Ack1, reduces tyrosine phosphorylation of focal adhesion kinase and p130Cas and disrupts the Crk·p130Cas complex. We further determined that suppression of expression of Cdc42, Ack1, p130Cas, or Crk reduces Rac1 activation and cell motility in Dbs-expressing cells to a level comparable with that in vector cells. Therefore, a cascade of activation of Cdc42 and Rac1 by Dbs through the Cdc42 effector Ack1 and the Crk·p130Cas complex is established. Suppression of the expression of endogenous Dbs reduces cell motility in both T47D cells and MDA-MB-231 cells, which correlates with the down-regulation of Cdc42 activity. This suggests that Dbs activates Cdc42 in these two human breast cancer cell lines and that the normal function of Dbs may be required to support cell movement.