Home page for Audrey Minden's Lab

Audrey Minden

Columbia University
Biological Sciences
813 Fairchild Bldg, M.C. 2460
New York, N.Y. 10027

E-mail agm24@columbia.edu

Return to Biology Home Page

In order to survive and develop normally, eukaryotic cells must be able to respond to a variety of extracellular stimuli and in some cases adapt to adverse conditions. Exposure to certain extracellular stimuli can trigger cell growth and division, while some stimuli can induce other responses such as cell differentiation, changes in cell shape, or even programmed cell death. Small GTPases of the Ras superfamily serve as molecular switches which mediate responses to many types of extracellular stimuli. Our lab is interested in understanding how small GTPases integrate signals from extracellular stimuli and in turn induce various cellular changes in mammalian cells. A major focus is the Rho family of GTPases, which are a subset of the Ras superfamily. Members of the Rho family, including Cdc42, Rac, and Rho, were first identified as proteins that have key roles in regulating the organization of the actin cytoskeleton and thereby control cell shape, motility, and adhesion. Subsequently, the Rho GTPases were found to have other functions including the regulation of cell proliferation and activation of signal transduction pathways that lead to changes in gene expression patterns. When improperly regulated, the Rho GTPases also play key roles in oncogenic transformation and tumor invasiveness. This most likely reflects the fact that changes in cell shape, adhesion, and motility, all play an important part in the oncogenic process. Furthermore, the Rho GTPases have been found to regulate important cell cycle regulatory genes which may cause cells to enter into the cell cycle and proliferate.

One area of research in our laboratory has been the identification of new molecular targets for the Rho GTPases. We have cloned genes for two new protein kinases that bind to and get activated by Cdc42. Both of these protein kinases are members of the PAK family of serine/threonine kinases. We have found that one of these kinases, PAK4, mediates cytoskeletal and cell shape changes in response to Cdc42 activation. Importantly, we have also found that a constitutively active mutant of PAK4 can induce anchorage oncogenic independent growth in fibroblasts. This may be due in part to effects this protein has on cell adhesion. This is especially intriguing because anchorage independent growth is a key step in the oncogenic process, and activated Cdc42 has been shown to have a similar effect. Furthermore, inhibition of PAK4 expression blocks oncogenic transformation by oncogenic Dbl, which is a potent oncogene and an activator of Cdc42. We predict that PAK4 is a key player in the signaling pathway by which Rho GTPases regulate oncogenic transformation. Currently we are investigating the substrates for PAK4 that may be directly responsible for its functions.

The second gene encodes another member of the PAK family, PAK5, which is also a target for Cdc42. Unlike PAK4, however, which is ubiquitously expressed, PAK5 is expressed mostly in the brain. The regulation of cytoskeletal organization and cell shape are particularly important in neuronal cells, especially during development when neurons extend axons and dendrites and connections are made with target cells. Consistent with this, we have found that activated PAK5 promotes neurite outgrowth when it is overexpressed in a neuroblastoma cell line. Experiments are currently underway in our lab to determine whether PAK5 is indeed an important player in the process of neurite outgrowth. This involves further studies in cell lines as well as a mouse knockout project in which we are generating mice which are deficient in the PAK5 gene. A similar mouse knockout project is underway for PAK4.

Finally, an important function of the Rho GTPases is the regulation of cell proliferation and cell cycle entry in both normal and transformed cells. In fact, the Rho GTPases are essential for proliferation and transformation by oncogenes such as the Ras oncogene. This may be due in part to their ability to control the expression of cell cycle regulatory proteins such as p21 and Cyclin D1. Ongoing projects in the lab involve deciphering the molecular mechanism by which Rho GTPases may regulate the expression of cell cycle regulatory proteins and thereby control cell proliferation.

Representative Publications:

Liberto, M., Cobrinik, D., Minden, A. (2002). Rho regulates. p21CIP1, cyclin D1, and checkpoint control in mammary epithelial cells. Oncogene. 21: 1590-1599.

Dan, C., Nath, N., Liberto, M. Minden, A. (2002). PAK5, a new brain specific kinase promotes neurite outgrowth in N1E-115 cells. Mol. Cell. Biol. 22: 567-577.

Dan, C., Kelley, A., Bernard, O., Minden, A. (2001) PAK4 induced cytoskeletal changes are mediated by LIMK1 and cofilin. J. Biol. Chem. 276: 32115-32121.

Cammarano, M.S., Minden, A. (2001) Dbl and the Rho GTPases activate NFKB by IKK dependent and IKK independent pathways. J. Biol. Chem. 276: 25876-25882.

Qu, J, Cammarano, M.S. Shi, Q., Ha, K.C. de Lanerolle, P., Minden, A. (2001). Activated PAK4 regulates cell adhesion and anchorage-independent growth. Mol. Cell. Biol. 21; 3523-3533.

Gnesutta, A., Qu, J., Minden, A. (2001). The Serine/Threonine Kinase PAK4 prevents caspase activation and protects cells from apoptosis. J. Biol. Chem. 276; 14414-14419.

Wang, S., Nath, N., Minden, A., Chellappan, S. (1999). Regulation of Rb and E2F by signal transduction cascades: Divergent effects of JNK1 and p38 kinases. EMBO J. 18: 1559-1570.

Minden, A., and Karin, M. (1998). The JNK Family of MAP Kinases: Regulation and Function. in Hormones and Signaling, B. W. O'Malley, Ed. Academic Press, pp. 209-233.

Abo. A., Qu, J., Cammarano, M., Dan, C., Fritsch, A., Minden, A. (1998). PAK4, a Novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of Filopodia. EMBO J 17: 6527-6540.

Minden, A., and Karin, M. (1997). Regulation and Function of the JNK subgroup of MAP Kinases. BBA: Reviews on Cancer. 1333: F85-F104

Kuroki, D.W., Minden, A., Sanchez, I., and Wattenberg, E.V. (1997) Regulation of a c-Jun amino terminal/stress-activated protein kinase cascade by a sodium-dependent signal transduction pathway. J. Biol Chem. 272:23905-23911.

Collins, L., Minden, A., Karin, M., and Brown J.H. (1996) G`12 stimulates c-Jun NH2- terminal kinase through the small G pro- teins Ras and Rac. J. Biol. Chem. 271:17349-17353Abstract

Lin, A., Minden, A., Martinetto, H., Claret, C.X., Lange-Carter, C., Mercurio, F., Johnson, G.L. & Karin, M. (1995) Identification of a dual specificity kinase that activates the Jun kinases and p38-Mpk2. Science 268:286-290. Abstract

Minden, A., Lin, A., Claret, F.X., Abo, A. & Karin, M. (1995) Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs. Cell 81:1147-1157. Abstract

Minden, A., Lin, A., Smeal, T., Cobb, M., and Karin, M. (1994) C-Jun N-terminal phosphorylation correlates with activation of the JNK subgroup but not the ERK subgroup of mitogen activated protein kinases. Mol. Cell. Biol. 14: 6683-6688Abstract

Minden, A., Lin, A., McMahon, M., Lange-Carter, C., Derijard, B., Davis, R.J., Johnson, G.L., and Karin, M. (1994) Differential activation of ERK and JNK mitogen activated protein kinases by Raf-1 and MEKK-1. Science 266:1719-1723. Abstract

Lab Members

Back to the top | MedLine List of Minden's Publications | Return to Biology Homepage