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Eukaryotic Cell, February 2003, p. 62-75, Vol. 2, No. 1
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.1.62-75.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Constitutively Active Protein Kinase A Disrupts Motility and Chemotaxis in Dictyostelium discoideum

Hui Zhang,1 Paul J. Heid,1 Deborah Wessels,1 Karla J. Daniels,1 Tien Pham,1 William F. Loomis,2 and David R. Soll1*

W. M. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242,1 Department of Biology, University of California, San Diego, La Jolla, California 920392

Received 27 August 2002/ Accepted 28 October 2002

The deletion of the gene for the regulatory subunit of protein kinase A (PKA) results in constitutively active PKA in the pkaR mutant. To investigate the role of PKA in the basic motile behavior and chemotaxis of Dictyostelium discoideum, pkaR mutant cells were subjected to computer-assisted two- and three-dimensional motion analysis. pkaR mutant cells crawled at only half the speed of wild-type cells in buffer, chemotaxed in spatial gradients of cyclic AMP (cAMP) but with reduced efficiency, were incapable of suppressing lateral pseudopods in the front of temporal waves of cAMP, a requirement for natural chemotaxis, did not exhibit the normal velocity surge in response to the front of a wave, and were incapable of chemotaxing toward an aggregation center in natural waves generated by wild-type cells that made up the majority of cells in mixed cultures. Many of the behavioral defects appeared to be the result of the constitutively ovoid shape of the pkaR mutant cells, which forced the dominant pseudopod off the substratum and to the top of the cell body. The behavioral abnormalities that pkaR mutant cells shared with regA mutant cells are discussed by considering the pathway ERK2 | RegA | [cAMP] -> PKA, which emanates from the front of a wave. The results demonstrate that cells must suppress PKA activity in order to elongate along a substratum, suppress lateral-pseudopod formation, and crawl and chemotax efficiently. The results also implicate PKA activation in dismantling cell polarity at the peak and in the back of a natural cAMP wave.

* Corresponding author. Mailing address: Department of Biological Science, BBE 302, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-1117. Fax: (319) 335-2772. E-mail: david-soll{at}uiowa.edu.

Eukaryotic Cell, February 2003, p. 62-75, Vol. 2, No. 1
1535-9778/03/$08.00+0     DOI: 10.1128/EC.2.1.62-75.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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