Pharmacological Sciences

Regulation of exocytosis, cell shape, and mitochondrial fusion by lipid signaling pathways

Our laboratory examines lipid signals that alter cell morphology, regulate mitochondrial fusion, and trigger subcellular trafficking of membrane vesicles during regulated exocytosis and endocytosis. Disease-related topics include neurodegenerative disease (Charcot-Marie Tooth syndrome), diabetes, cancer, and impaired phagocytic immune responses to pathogens.

The specific topic constitutes study of signaling pathways mediated by members of the superfamiles of enzymes known as Phospholipase D (PLD) and PI4P Kinase. Classic PLD is activated by a wide variety of agonists that signal through G-protein coupled or tyrosine kinase receptors. PLD has varied cellular roles including facilitating membrane vesicle trafficking and fusion of the vesicles into the plasma membrane during regulated exocytosis, and reorganizing the actin cytoskeleton. We are currently investigating roles for PLD and PI4P lipid kinase in actin cytoskeleton reorganization, myoblast differentiation and fusion, Glut-4 glucose transporter translocation, phagocytosis, and regulated exocytosis (insulin and histamine release). In addition, we are exploring a novel member of the PLD superfamily that localizes to mitochondria and regulates fusion, and the mechanisms through which diminished rates of fusion cause neurodegenerative disease.

Lab Members:

Former Lab Members:

Selected Publications:

Su, W. and Frohman, M.A. (2007)Phospholipase D. In: Lipid-mediated Signal Transduction (Rosenberger, T. and Murphy, E.J., Editors), CRC Press Methods in Signal Transduction, in press.

Scotto-Lavino, E., Du, G., and Frohman, M.A. Using RACE to clone cDNA ends. (2006) Nature Protocols, in press.

Vicogne, J, Vollenweider, D, Smith, J, Huang, P, Frohman, MA and Pessin, J (2006) Asymmetric phospholipid distribution drives in vitro reconstituted SNARE-dependent membrane fusion. Proc Natl Acad Sci, in press.

Choi, SY, Huang, P, Jenkins, GM, Chan, DC, Schiller, J, and Frohman, MA (2006) A common signaling lipid requirement for Mfn-mediated mitochondrial
fusion and SNARE-regulated exocytosis. Nature Cell Biol, in press.

Choi SY, Gonzalvez F, Jenkins GM, Slomianny C, Chretien D, Arnoult D, Petit PX, Frohman MA (2006). Cardiolipin deficiency releases cytochrome c from
the inner mitochondrial membrane and accelerates stimuli-elicited apoptosis. Cell Death Differ, in press.

Cai D, Netzer WJ, Zhong M, Lin Y, Du G, Frohman M, Foster DA, Sisodia SS, Xu H, Gorelick FS, Greengard P (2006). Presenilin-1 uses phospholipase D1 as a negative regulator of beta-amyloid formation. Proc Natl Acad Sci U S A. 103: 1941-6.

Corrotte M, Chasserot-Golaz S, Huang P, Du G, Ktistakis NT, Frohman MA, Vitale N, Bader MF, Grant NJ (2006). Dynamics and function of phospholipase D and phosphatidic acid during phagocytosis. Traffic, 7(3):365-77.

Suzuki T, Kanai Y, Hara T, Sasaki J, Sasaki T, Kohara M, Maehama T, Taya C, Shitara H, Yonekawa H, Frohman MA, Yokozeki T, Kanaho Y (2006). Crucial role of the small GTPase ARF6 in hepatic cord formation during liver development. Mol Cell Biol. 26: 6149-56.

Huang P, Jiang Z, Teng S, Wong YC, Frohman MA, Chung SK, Chung SS (2006). Synergism between phospholipase D2 and sorbitol accumulation in diabetic cataract formation through modulation of Na,K-ATPase activity and osmotic stress. Exp Eye Res83(4): 939-948.

Andersson L, Bostrm P, Ericson J, Rutberg M, Magnusson B, Marchesan D, Ruiz M, Asp L, Huang P, Frohman MA, Born J, Olofsson SO (2006). PLD1 and
ERK2 regulate cytosolic lipid droplet formation. J Cell Sci. 119: 2246-57.

Jenkins GM, Frohman MA. (2005)Phospholipase D: a lipid centric review. Cell Mol Life Sci. 62(19-20):2305-16.

Xiao N, Du G, Frohman MA. (2005)Peroxiredoxin II functions as a signal terminator for H2O2-activated phospholipase D1. FEBS J. 272(15):3929-37.

LaLonde, M.M., Janssen, H., Rosenbaum, E., Choi, S.-Y., Gergen, J.P., Colley, N.J., Stark, W.S., and Frohman, M.A.(2005) Regulation of phototransduction responsiveness and retinal degeneration by a phospholipase D-generated signaling lipid. J. Cell  Biol. 169(3):471-9.

Huang, P., Altshuller, Y.M., Hou, J.C., Pessin, J.E., and Frohman, M.A. (2005) Insulin-stimulated plasma membrane fusion of Glut4 glucose transporter-containing vesicles is regulated by Phospholipase D1. Mol. Biol. Cell,. 16(6):2614-23.

Hughes WE, Elgundi Z, Huang P, Frohman MA, Biden TJ (2004). Phospholipase D1 regulates secretagogue-stimulated insulin release in pancreatic beta-cells. J Biol Chem. 279 27534-41.

Watanabe H, Yamazaki M, Miyazaki H, Arikawa C, Itoh K, Sasaki T, Maehama T, Frohman M.A., Kanaho Y. (2004)Phospholipase D2 functions as a downstream signaling molecule of MAP kinase pathway in L1-stimulated neurite outgrowth of cerebellar granule neurons. J Neurochem. 89(1):142-51.

Zhang Y, Huang P, Du G, Kanaho Y, Frohman M.A., Tsirka SE. (2004) Increased expression of two phospholipase D isoforms during experimentally induced hippocampal mossy fiber outgrowth. Glia. 46(1):74-83.

Du G, Huang P, Liang BT, Frohman M.A. (2004) Phospholipase D2 Localizes to the Plasma Membrane and Regulates Angiotensin II Receptor Endocytosis. Mol Biol Cell. 2004 Mar;15(3):1024-30.

Zheng XL, Gui Y, Du G, Frohman M.A., Peng DQ. (2004) Calphostin-C induction of vascular smooth muscle cell apoptosis proceeds through phospholipase D and microtubule inhibition. J Biol Chem. 279(8):7112-8.

Fang Y, Park IH, Wu AL, Du G, Huang P, Frohman M.A., Walker SJ, Brown HA, Chen J. (2003) PLD1 Regulates mTOR Signaling and Mediates Cdc42 Activation of S6K1. Curr Biol. 13:2037-44.

Huang, P. and Frohman, M.A. (2003) Phospholipase D and Glut-4 trafficking. Diabetes Metab Res Rev. 19:456-63

Ballou LM, Jiang YP, Du G, Frohman M.A., Lin RZ. (2003) Ca(2+)- and phospholipase D-dependent and -independent pathways activate mTOR signaling. FEBS Lett. 550:51-6.

Du, G., Altshuller, Y.M., Vitale, N., Huang, P., Chasserot-Golaz, S., Morris, A.J., Bader, M.-F., and Frohman, M.A. (2003) Regulation of Phospholipase D1 subcellular cycling through coordination of multiple membrane association motifs. J.Cell Biol. 162(2):305-15

Du, G., Morris, A.J., Sciorra, V.A., and Frohman, M.A. (2002) G-protein coupled receptor regulation of Phospholipase D. In : G-protein Pathways. Methods in Enzymology (Hildebrandt, J.D. and Iyengar, R., Editor). 345:265-274.

Yamazaki, M., Miyazaki, H., Watanabe, H., Maehama, T., Frohman, M.A., and Kanaho, Y. (2002) Phosphatidylinositol 4-phosphate 5-kinase couples ROCK to neurite remodeling. J. Biol. Chem, 277:17226-17230.

Choi., W.S., Kim, Y.M., Combs, C., Frohman, M.A., and Beaven, M.A. (2002) Phospholipase D1 and 2 Regulate Different Phases of Exocytosis in Mast Cells. J. Immunology 168:5682-9.

Vitale, N., Chasserot-Golaz, S., Bailly, Y., Morinaga, N., Frohman, M.A., and Bader, M.-F. (2002) Calcium-regulated exocytosis of dense core vesicles requires the activation of ARF6 by ARNOat the plasma membrane. J. Cell Biol. 159:79-89.

Denmat-Ouisse LA, Phebidias C, Honkavaara P, Robin P, Geny B, Min do S, Bourgoin S, Frohman MA, Raymond MN. (2001) Regulation of constitutive protein transit by phospholipase D in HT29-cl19A cells. J. Biol.Chem. 276:48840-6.

Vitale, N., Caumont-Primus, A.-S., Chasserot-Golaz, S., Du, G., Wu, S., Sciorra, V.A., Morris, A.J., Frohman, M.A. and Bader, M.-F. (2001) Phospholipase D1 : a key factor for the exocytotic machinery in neuroendocrine cells. EMBO J., 20(10):2424-2434.

Freyberg, Z., Siddhanta, A., Sweeney, D., Bourgoin, S., Frohman, M.A., and Shields, D. (2001) Intracellular Localization of Phospholipase D1 in Mammalian Cells. Mol. Biol. Cell., 12(4):943-955.

Humeau, Y., Vitale, N., Chasserot-Golaz, S., Dupont, J.-L., Du, G., Frohman, M.A., Bader, M.-F. and Poulain, P. (2001) A role for phospholipase D1 in neurotransmitter release. Proc. Natl. Acad. Sci. U.S.A.. 98:15300-5.

Du, G., Altshuller, Y.M., Kim., Y., Han, J.M., Ryu, S.H., Morris, A.J., and Frohman, M.A. (2000) Dual requirement for Rho and protein kinase C in direct activation of phospholipase D1 through G-protein coupled receptor signaling. Mol. Biol. Cell., 11:4359-4368.

Slaaby, R., Du, G., Altshuller, Y. M., Frohman, M.A., and Seedorf, K. (2000) PKCa and PLCg are involved in the activation of PLD1 and PLD2 upon insulin stimulation. Biochem. J., 351:613-619.

Zhang, Y., Redina, O., Altshuller, Y.M., Yamazaki, M., Ramos, J., Chneiweis, H., Kanaho, Y., and Frohman, M.A. (2000) Regulation of PLD1 and PLD2 expression levels by Pea-15, a novel PLD-interacting protein.  J. Biol. Chem., 275: 35224-35232.

Park, J.B., Kim, J.H., Kim, J.H., Kim, J, Yoo, J.-S.,Suh, P.-G., Du, G., Frohman, M.A., and Ryu, S.H. (2000) Cardiac phospholipase D2 localizes to sarcolemmal membranes and is inhibited by alpha-actinin in an ADP-ribosylation factor-reversible manner. J. Biol. Chem., 275: 21295-21301.

Lee, C., Kim, S.R., Chung, J.-K., Frohman, M.A., Kilimann, M.W., Rhee, S.G. (2000) Inhibition of Phospholipase D by Amphiphysins. J. Biol. Chem., 275:
18751-18758.

Wu, Y.P., Lu, W., Sung, T-C., Frohman, M.A., Milev, P., Bugge, T.H., Degen, J.L.,Margolis, R.U., and Tsirka, S.E. (2000) The tPA/plasmin extracellular proteolytic system contributes to hippocampal mossy fiber reorganization through a novel proteoglycan substrate. J. Cell Biol., 148:1295-304.

Honda, A., Nogami, M., Yokozeki, T., Yamazaki, M., Nakamura, H., Watanabe, H., Kawamoto, K., Nakayama, K., Morris, A.J., Frohman, M.A., and Kanaho, Y. (1999) Phosphatidylinositol 4-phosphate 5-kinasea is a Downstream
Effector of the Small G Protein ARF6 in Membrane Ruffle Formation. Cell, 99:521-532.

Sciorra, V.A, Rudge, S., Prestwich, G.D., Frohman, M.A., Engebrecht, J., and Morris, A.J. (1999) Identification of a phosphoinositide binding motif that mediates activation of mammalian and
yeast Phospholipase D isoenzymes. EMBO J., 18: 5911-5921.

Zhang, Y, Altshuller, Y.A., Morris, A.J., and Frohman, M.A. (1999) Loss of Receptor Regulation by a Phospholipase D1 mutant unresponsive to Protein Kinase C.  EMBO J., 18:6339-6348.

Yamazaki, M., Zhang, Y., Watanabe, H., Yokozeki, T., Ohno, S., Kaibuchi, K., Shibata, H., Mukai, H., Ono, Y., Frohman, M.A., and Kanaho, Y. (1999) Identification of the RhoA-binding domain of Phospholipase D1. J. Biol. Chem., 
274:6035-6038.

Rizzo, M.A., Shome, K., Vasudevan, C., Stolz, D.B., Sung, T.-C., Frohman, M.A., Watkins, S.C., and Romero, G. (1999)Phospholipase D and its product, phosphatidic acid, mediate agonist-dependent Raf-1 translocation to the plasma membrane and the activation of the MAP kinase pathway. J. Biol. Chem., 274: 1131-1139.

Sung, T.-C., Altshuller, Y.M., Morris, A.J. and Frohman, M.A. (1999) Molecular Analysis of Mammalian Phospholipase D2. J. Biol. Chem. 274:494-502.

Sung, T.-C., Zhang, Y., Morris, A.J. and Frohman, M.A. (1999) Structural Analysis of Human Phospholipase D1. J. Biol. Chem., 274:3659-3666.

Sung, T.-C., Roper, K., Zhang, Y., Rudge, S., Temel, R., Hammond, S.M., Morris, A.J., Moss, B., Engebrecht, J., and Frohman, M.A. (1997) Mutagenesis of Phospholipase D defines a superfamily including a trans-Golgi viral protein required for poxvirus pathogenicity. EMBO J. 16:4519-4530.

Chong, J.A., Tapia-Ramírez, J., Kim, S., Toledo-Aral, J.J., Zheng, Y., Boutros, M.C., Altshuller, Y.M., Frohman, M.A., Kraner, S.D., and Mandel, G. (1995) REST:  A Mammalian Silencer Protein that Restricts Sodium Channel Gene Expression to Neurons. Cell 80:949-957.