RasC plays a role in transduction of temporal gradient information in the cyclic-AMP wave of Dictyostelium discoideum.
To define the role that RasC plays in motility and chemotaxis, the behavior of a rasC null mutant, rasC-, in buffer and in response to the individual spatial, temporal, and concentration components of a natural cyclic AMP (cAMP) wave was analyzed by using computer-assisted two-dimensional and three-dimensional motion analysis systems. These quantitative studies revealed that rasC- cells translocate at the same velocity and exhibit chemotaxis up spatial gradients of cAMP with the same efficiency as control cells. However, rasC- cells exhibit defects in maintaining anterior-posterior polarity along the substratum and a single anterior pseudopod when translocating in buffer in the absence of an attractant. rasC- cells also exhibit defects in their responses to both the increasing and decreasing temporal gradients of cAMP in the front and the back of a wave. These defects result in the inability of rasC- cells to exhibit chemotaxis in a natural wave of cAMP. The inability to respond normally to temporal gradients of cAMP results in defects in the organization of the cytoskeleton, most notably in the failure of both F actin and myosin II to exit the cortex in response to the decreasing temporal gradient of cAMP in the back of the wave. While the behavioral defect in the front of the wave is similar to that of the myoA-/myoF- myosin I double mutant, the behavioral and cytoskeletal defects in the back of the wave are similar to those of the S13A myosin II regulatory light-chain phosphorylation mutant. Expression array data support the premise that the behavioral defects exhibited by the rasC- mutant are the immediate result of the absence of RasC function.
Wessels
Deborah
D
W. M. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, The University of Iowa, Iowa City, IA 52242, USA.
Brincks
Rebecca
R
Kuhl
Spencer
S
Stepanovic
Vesna
V
Daniels
Karla J
KJ
Weeks
Gerald
G
Lim
Chinten J
CJ
Spiegelman
George
G
Fuller
Danny
D
Iranfar
Negin
N
Loomis
William F
WF
Soll
David R
DR
D000818:Animals; D002465:Cell Movement / Q000502:physiology*; D002633:Chemotaxis / Q000502:physiology*; D000242:Cyclic AMP / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D004023:Dictyostelium / Q000235:genetics / Q000502:physiology*; D009154:Mutation / Q000235:genetics; D024462:Myosin Type II / Q000378:metabolism; D009218:Myosins / Q000378:metabolism; D010766:Phosphorylation; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011554:Pseudopodia / Q000378:metabolism; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*
D015800:Protozoan Proteins; D000242:Cyclic AMP; D024462:Myosin Type II; D009218:Myosins; D018631:ras Proteins