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Journal of Cell Science 115, 1733-1747 (2002)
© 2002 The Company of Biologists Limited

Research Article

Phosphorylation of the myosin regulatory light chain plays a role in motility and polarity during Dictyostelium chemotaxis

Hui Zhang1, Deborah Wessels1, Petra Fey2, Karla Daniels1, Rex L. Chisholm2 and David R. Soll1,*

1 Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242, USA
2 Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA

* Author for correspondence (e-mail: david-soll{at}uiowa.edu )

Accepted 29 January 2002

The myosin regulatory light chain (RLC) of Dictyostelium discoideum is phosphorylated at a single serine site in response to chemoattractant. To investigate the role of the phosphorylation of RLC in both motility and chemotaxis, mutants were generated in which the single phosphorylatable serine was replaced with a nonphosphorylatable alanine. Several independent clones expressing the mutant RLC in the RLC null mutant, mlcR-, were obtained. These S13A mutants were subjected to high resolution computer-assisted motion analysis to assess the basic motile behavior of cells in the absence of a chemotatic signal, and the chemotactic responsiveness of cells to the spatial, temporal and concentration components of natural cAMP waves. In the absence of a cAMP signal, mutant cells formed lateral pseudopods less frequently and crawled faster than wild-type cells. In a spatial gradient of cAMP, mutant cells chemotaxed more efficiently than wild-type cells. In the front of simulated temporal and natural waves of cAMP, mutant cells responded normally by suppressing lateral pseudopod formation. However, unlike wild-type cells, mutant cells did not lose cellular polarity at the peak and in the back of either wave. Since depolarization at the peak and in the descending phase of the natural wave is necessary for efficient chemotaxis, this deficiency resulted in a decrease in the capacity of S13A mutant cells to track natural cAMP waves relayed by wild-type cells, and in the fragmentation of streams late in mutant cell aggregation. These results reveal a regulatory pathway induced by the peak and back of the chemotactic wave that alters RLC phosphorylation and leads to cellular depolarization. We suggest that depolarization requires myosin II rearrangement in the cortex facilitated by RLC phosphorylation, which increases myosin motor function.

Key words: Myosin light chain, Myosin phosphorylation, Cell motility, Chemotaxis, Dictyostelium discoideum




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© The Company of Biologists Ltd 2002