Characterization of AND-34 function and signaling.
AND-34 is a member of a novel family of proteins (NSP1, NSP2, and NSP3) that have an amino-terminal SH2 domain but bind by a carboxy-terminal GEF (Cdc25)-like domain to the carboxy-terminus of the focal adhesion adapter protein p130Cas. Direct GEF activity of AND-34 toward Ras subfamily members has not been demonstrated with purified protein. Overexpression of AND-34 in epithelial breast cancer cells leads to activation of Rac and Cdc42 by a PI3K-dependent mechanism. This chapter will describe the techniques we used to examine AND-34-induced Rac, Cdc42, Akt, and PAK1 activation in human breast cancer cell lines and in murine lymphoid cell lines. In addition, we summarize techniques used to determine that AND-34 overexpression does not activate R-Ras in MCF-7 cells.
Felekkis
Kyriacos
K
Department of Pathology, Boston University School of Medicine, Boston, Massachusetts, USA.
Quilliam
Lawrence A
LA
Lerner
Adam
A
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001402:B-Lymphocytes; D045744:Cell Line, Tumor; D020662:Guanine Nucleotide Exchange Factors / Q000502:physiology*; D006801:Humans; D051379:Mice; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D054462:p21-Activated Kinases / Q000378:metabolism; D018631:ras Proteins / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; C120867:Bcar3 protein, mouse; D020662:Guanine Nucleotide Exchange Factors; D051057:Proto-Oncogene Proteins c-akt; D054462:p21-Activated Kinases; C085929:RRAS protein, human; D020764:cdc42 GTP-Binding Protein; D018631:ras Proteins