Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Cell Biol. 147 (3): 559-576

Copyright © 1999 by the Rockefeller University Press.

Original Article

Paka, a Putative Pak Family Member, Is Required for Cytokinesis and the Regulation of the Cytoskeleton in Dictyostelium discoideum Cells during Chemotaxis

Chang Y. Chunga, and Richard A. Firtela

a Department of Biology, Center for Molecular Genetics, University of California San Diego, La Jolla, California 92093-0634
Center for Molecular Genetics, Room 225, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0634.(858) 534-7073(858) 534-2788


Abstract: We have identified a Dictyostelium discoideum gene encoding a serine/threonine kinase, PAKa, a putative member of the Ste20/PAK family of p21-activated kinases, with a kinase domain and a long NH2-terminal regulatory domain containing an acidic segment, a polyproline domain, and a CRIB domain. PAKa colocalizes with myosin II to the cleavage furrow of dividing cells and the posterior of polarized, chemotaxing cells via its NH2-terminal domain. paka null cells are defective in completing cytokinesis in suspension. PAKa is also required for maintaining the direction of cell movement, suppressing lateral pseudopod extension, and proper retraction of the posterior of chemotaxing cells. paka null cells are defective in myosin II assembly, as the myosin II cap in the posterior of chemotaxing cells and myosin II assembly into cytoskeleton upon cAMP stimulation are absent in these cells, while constitutively active PAKa leads to an upregulation of myosin II assembly. PAKa kinase activity against histone 2B is transiently stimulated and PAKa incorporates into the cytoskeleton with kinetics similar to those of myosin II assembly in response to chemoattractant signaling. However, PAKa does not phosphorylate myosin II. We suggest that PAKa is a major regulator of myosin II assembly, but does so by negatively regulating myosin II heavy chain kinase.

Key Words: Dictyostelium discoideum • PAK • myosin II • chemotaxis • Rac/Cdc42

1.used in this paper: GFP, green fluorescent protein; H2B, histone 2B; MIHCK, myosin I heavy chain kinase; myrPAKa, a PAKa carrying the NH2-terminal myristoylation signal from Src; PAKaCA, a putative constitutively active PAKa containing only the kinase domain; PAKaDN, kinase dead, putative dominant, negative mutant of PAKa; PAKaG+K, a PAKa mutant containing the CRIB and kinase domains

A Cdc42- and Rac-interactive binding (CRIB) domain mediates functions of coronin.
K. Swaminathan, A. Muller-Taubenberger, J. Faix, F. Rivero, and A. A. Noegel (2014)
PNAS 111, E25-E33
   Abstract »    Full Text »    PDF »
PakD, a Putative p21-Activated Protein Kinase in Dictyostelium discoideum, Regulates Actin.
M. Garcia, S. Ray, I. Brown, J. Irom, and D. Brazill (2014)
Eukaryot. Cell 13, 119-126
   Abstract »    Full Text »    PDF »
Daydreamer, a Ras effector and GSK-3 substrate, is important for directional sensing and cell motility.
V. Kolsch, Z. Shen, S. Lee, K. Plak, P. Lotfi, J. Chang, P. G. Charest, J. L. Romero, T. J. Jeon, A. Kortholt, et al. (2013)
Mol. Biol. Cell 24, 100-114
   Abstract »    Full Text »    PDF »
Integrins in Cell Migration.
A. Huttenlocher and A. R. Horwitz (2011)
Cold Spring Harb Perspect Biol 3, a005074
   Abstract »    Full Text »    PDF »
Disruption of PKB signaling restores polarity to cells lacking tumor suppressor PTEN.
M. Tang, M. Iijima, Y. Kamimura, L. Chen, Y. Long, and P. Devreotes (2011)
Mol. Biol. Cell 22, 437-447
   Abstract »    Full Text »    PDF »
BMP2 induction of actin cytoskeleton reorganization and cell migration requires PI3-kinase and Cdc42 activity.
C. Gamell, N. Osses, R. Bartrons, T. Ruckle, M. Camps, J. L. Rosa, and F. Ventura (2008)
J. Cell Sci. 121, 3960-3970
   Abstract »    Full Text »    PDF »
Linking Ras to myosin function: RasGEF Q, a Dictyostelium exchange factor for RasB, affects myosin II functions.
S. Mondal, D. Bakthavatsalam, P. Steimle, B. Gassen, F. Rivero, and A. A. Noegel (2008)
J. Cell Biol. 181, 747-760
   Abstract »    Full Text »    PDF »
A Protein with Similarity to PTEN Regulates Aggregation Territory Size by Decreasing Cyclic AMP Pulse Size during Dictyostelium discoideum Development.
Y. Tang and R. H. Gomer (2008)
Eukaryot. Cell 7, 1758-1770
   Abstract »    Full Text »    PDF »
Combining experiments and modelling to understand size regulation in Dictyostelium discoideum.
W. Jang and R. H Gomer (2008)
J R Soc Interface 5, S49-S58
   Abstract »    Full Text »    PDF »
The regulation of cell motility and chemotaxis by phospholipid signaling.
V. Kolsch, P. G. Charest, and R. A. Firtel (2008)
J. Cell Sci. 121, 551-559
   Abstract »    Full Text »    PDF »
The Ste20-like kinase SvkA of Dictyostelium discoideum is essential for late stages of cytokinesis.
M. Rohlfs, R. Arasada, P. Batsios, J. Janzen, and M. Schleicher (2007)
J. Cell Sci. 120, 4345-4354
   Abstract »    Full Text »    PDF »
Regulation of Rap1 activity by RapGAP1 controls cell adhesion at the front of chemotaxing cells.
T. J. Jeon, D.-J. Lee, S. Lee, G. Weeks, and R. A. Firtel (2007)
J. Cell Biol. 179, 833-843
   Abstract »    Full Text »    PDF »
WASP-interacting Protein Is Important for Actin Filament Elongation and Prompt Pseudopod Formation in Response to a Dynamic Chemoattractant Gradient.
S. A. Myers, L. R. Leeper, and C. Y. Chung (2006)
Mol. Biol. Cell 17, 4564-4575
   Abstract »    Full Text »    PDF »
Guanylyl Cyclase Protein and cGMP Product Independently Control Front and Back of Chemotaxing Dictyostelium Cells.
D. M. Veltman and P. J.M. Van Haastert (2006)
Mol. Biol. Cell 17, 3921-3929
   Abstract »    Full Text »    PDF »
Loss of SMEK, a Novel, Conserved Protein, Suppresses mek1 Null Cell Polarity, Chemotaxis, and Gene Expression Defects.
M. C. Mendoza, F. Du, N. Iranfar, N. Tang, H. Ma, W. F. Loomis, and R. A. Firtel (2005)
Mol. Cell. Biol. 25, 7839-7853
   Abstract »    Full Text »    PDF »
Multiple Myosin II Heavy Chain Kinases: Roles in Filament Assembly Control and Proper Cytokinesis in Dictyostelium.
S. Yumura, M. Yoshida, V. Betapudi, L. S. Licate, Y. Iwadate, A. Nagasaki, T. Q.P. Uyeda, and T. T. Egelhoff (2005)
Mol. Biol. Cell 16, 4256-4266
   Abstract »    Full Text »    PDF »
Cdc42 controls the polarity of the actin and microtubule cytoskeletons through two distinct signal transduction pathways.
J. Cau and A. Hall (2005)
J. Cell Sci. 118, 2579-2587
   Abstract »    Full Text »    PDF »
RasGEF-containing proteins GbpC and GbpD have differential effects on cell polarity and chemotaxis in Dictyostelium.
L. Bosgraaf, A. Waijer, R. Engel, A. J. W. G. Visser, D. Wessels, D. Soll, and P. J. M. van Haastert (2005)
J. Cell Sci. 118, 1899-1910
   Abstract »    Full Text »    PDF »
A Dictyostelium Homologue of WASP Is Required for Polarized F-Actin Assembly during Chemotaxis.
S. A. Myers, J. W. Han, Y. Lee, R. A. Firtel, and C. Y. Chung (2005)
Mol. Biol. Cell 16, 2191-2206
   Abstract »    Full Text »    PDF »
Cellular Distribution and Functions of Wild-Type and Constitutively Activated Dictyostelium PakB.
M. de la Roche, A. Mahasneh, S.-F. Lee, F. Rivero, and G. P. Cote (2005)
Mol. Biol. Cell 16, 238-247
   Abstract »    Full Text »    PDF »
Dictyostelium PAKc Is Required for Proper Chemotaxis.
S. Lee, F. Rivero, K. C. Park, E. Huang, S. Funamoto, and R. A. Firtel (2004)
Mol. Biol. Cell 15, 5456-5469
   Abstract »    Full Text »    PDF »
Localized Ras signaling at the leading edge regulates PI3K, cell polarity, and directional cell movement.
A. T. Sasaki, C. Chun, K. Takeda, and R. A. Firtel (2004)
J. Cell Biol. 167, 505-518
   Abstract »    Full Text »    PDF »
Moving Forward: Mechanisms of Chemoattractant Gradient Sensing.
J. Franca-Koh and P. N. Devreotes (2004)
Physiology 19, 300-308
   Abstract »    Full Text »    PDF »
A Cell Number Counting Factor Regulates Akt/Protein Kinase B To Regulate Dictyostelium discoideum Group Size.
T. Gao, D. Knecht, L. Tang, R. D. Hatton, and R. H. Gomer (2004)
Eukaryot. Cell 3, 1176-1184
   Abstract »    Full Text »    PDF »
The genetics of Pak.
C. Hofmann, M. Shepelev, and J. Chernoff (2004)
J. Cell Sci. 117, 4343-4354
   Abstract »    Full Text »    PDF »
The Cyclase-associated Protein CAP as Regulator of Cell Polarity and cAMP Signaling in Dictyostelium.
A. A. Noegel, R. Blau-Wasser, H. Sultana, R. Muller, L. Israel, M. Schleicher, H. Patel, and C. J. Weijer (2004)
Mol. Biol. Cell 15, 934-945
   Abstract »    Full Text »    PDF »
Chemotaxis: signalling modules join hands at front and tail.
M. Postma, L. Bosgraaf, H. M. Loovers, and P. J. M. Van Haastert (2004)
EMBO Rep. 5, 35-40
   Abstract »    Full Text »    PDF »
Shear flow-induced motility of Dictyostelium discoideum cells on solid substrate.
E. Decave, D. Rieu, J. Dalous, S. Fache, Y. Brechet, B. Fourcade, M. Satre, and F. Bruckert (2003)
J. Cell Sci. 116, 4331-4343
   Abstract »    Full Text »    PDF »
Dictyostelium Stress-activated Protein Kinase {alpha}, a Novel Stress-activated Mitogen-activated Protein Kinase Kinase Kinase-like Kinase, Is Important for the Proper Regulation of the Cytoskeleton.
B. Sun, H. Ma, and R. A. Firtel (2003)
Mol. Biol. Cell 14, 4526-4540
   Abstract »    Full Text »    PDF »
Leading the way: directional sensing through phosphatidylinositol 3-kinase and other signaling pathways.
S. Merlot and R. A. Firtel (2003)
J. Cell Sci. 116, 3471-3478
   Abstract »    Full Text »    PDF »
A Regulator of G Protein Signaling-containing Kinase Is Important for Chemotaxis and Multicellular Development in Dictyostelium.
B. Sun and R. A. Firtel (2003)
Mol. Biol. Cell 14, 1727-1743
   Abstract »    Full Text »    PDF »
Rac and Cdc42 play distinct roles in regulating PI(3,4,5)P3 and polarity during neutrophil chemotaxis.
S. Srinivasan, F. Wang, S. Glavas, A. Ott, F. Hofmann, K. Aktories, D. Kalman, and H. R. Bourne (2003)
J. Cell Biol. 160, 375-385
   Abstract »    Full Text »    PDF »
EhPAK, a member of the p21-activated kinase family, is involved in the control of Entamoeba histolytica migration and phagocytosis.
E. Labruyere, C. Zimmer, V. Galy, J.-C. Olivo-Marin, and N. Guillen (2003)
J. Cell Sci. 116, 61-71
   Abstract »    Full Text »    PDF »
Intracellular localization and dynamics of myosin-II and myosin-IC in live Acanthamoeba by transient transfection of EGFP fusion proteins.
H.-H. Kong and T. D. Pollard (2002)
J. Cell Sci. 115, 4993-5002
   Abstract »    Full Text »    PDF »
Requirement of a Vasodilator-stimulated Phosphoprotein Family Member for Cell Adhesion, the Formation of Filopodia, and Chemotaxis in Dictyostelium.
Y.-H. Han, C. Y. Chung, D. Wessels, S. Stephens, M. A. Titus, D. R. Soll, and R. A. Firtel (2002)
J. Biol. Chem. 277, 49877-49887
   Abstract »    Full Text »    PDF »
PAK4 is activated via PI3K in HGF-stimulated epithelial cells.
C. M. Wells, A. Abo, and A. J. Ridley (2002)
J. Cell Sci. 115, 3947-3956
   Abstract »    Full Text »    PDF »
Cytokinesis in Eukaryotes.
D. A. Guertin, S. Trautmann, and D. McCollum (2002)
Microbiol. Mol. Biol. Rev. 66, 155-178
   Abstract »    Full Text »    PDF »
Phosphorylation of the myosin regulatory light chain plays a role in motility and polarity during Dictyostelium chemotaxis.
H. Zhang, D. Wessels, P. Fey, K. Daniels, R. L. Chisholm, and D. R. Soll (2002)
J. Cell Sci. 115, 1733-1747
   Abstract »    Full Text »    PDF »
A cell number-counting factor regulates the cytoskeleton and cell motility in Dictyostelium.
L. Tang, T. Gao, C. McCollum, W. Jang, M. G. Vicker, R. R. Ammann, and R. H. Gomer (2002)
PNAS 99, 1371-1376
   Abstract »    Full Text »    PDF »
Role of Phosphatidylinositol 3' Kinase and a Downstream Pleckstrin Homology Domain-Containing Protein in Controlling Chemotaxis inDictyostelium.
S. Funamoto, K. Milan, R. Meili, and R. A. Firtel (2001)
J. Cell Biol. 153, 795-810
   Abstract »    Full Text »    PDF »
An SH2-domain-containing kinase negatively regulates the phosphatidylinositol-3 kinase pathway.
J. Moniakis, S. Funamoto, M. Fukuzawa, J. Meisenhelder, T. Araki, T. Abe, R. Meili, T. Hunter, J. Williams, and R. A. Firtel (2001)
Genes & Dev. 15, 687-698
   Abstract »    Full Text »
Tortoise, a Novel Mitochondrial Protein, Is Required for Directional Responses of Dictyostelium in Chemotactic Gradients.
S. van Es, D. Wessels, D. R. Soll, J. Borleis, and P. N. Devreotes (2001)
J. Cell Biol. 152, 621-632
   Abstract »    Full Text »    PDF »
Temporal and Spatial Distribution of Activated Pak1 in Fibroblasts.
M. A. Sells, A. Pfaff, and J. Chernoff (2000)
J. Cell Biol. 151, 1449-1458
   Abstract »    Full Text »    PDF »
The Internal Phosphodiesterase RegA Is Essential for the Suppression of Lateral Pseudopods during Dictyostelium Chemotaxis.
D. J. Wessels, H. Zhang, J. Reynolds, K. Daniels, P. Heid, S. Lu, A. Kuspa, G. Shaulsky, W. F. Loomis, and D. R. Soll (2000)
Mol. Biol. Cell 11, 2803-2820
   Abstract »    Full Text »
Role of Rac in controlling the actin cytoskeleton and chemotaxis in motile cells.
C. Y. Chung, S. Lee, C. Briscoe, C. Ellsworth, and R. A. Firtel (2000)
PNAS 97, 5225-5230
   Abstract »    Full Text »    PDF »
The Dictyostelium LIM Domain-containing Protein LIM2 Is Essential for Proper Chemotaxis and Morphogenesis.
S. Chien, C. Y. Chung, S. Sukumaran, N. Osborne, S. Lee, C. Ellsworth, J. G. McNally, and R. A. Firtel (2000)
Mol. Biol. Cell 11, 1275-1291
   Abstract »    Full Text »
Untying the Gordian Knot of Cytokinesis: Role of Small G Proteins and Their Regulators.
S. N. Prokopenko, R. Saint, and H. J. Bellen (2000)
J. Cell Biol. 148, 843-848
   Full Text »    PDF »
The actin cytoskeleton of Dictyostelium: a story told by mutants.
A. A. Noegel and M. Schleicher (2000)
J. Cell Sci. 113, 759-766
   Abstract »    PDF »
A cell number-counting factor regulates the cytoskeleton and cell motility in Dictyostelium.
L. Tang, T. Gao, C. McCollum, W. Jang, M. G. Vicker, R. R. Ammann, and R. H. Gomer (2002)
PNAS 99, 1371-1376
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882