Phospholipase D1 regulates cell migration in a lipase activity-independent manner.
Cell migration, a complex biological process, requires dynamic cytoskeletal remodeling. Phospholipase D (PLD) generates phosphatidic acid, a lipid second messenger. Although PLD activity has been proposed to play a role in cytoskeletal rearrangement, the manner in which PLD participates in the rearrangement process remains obscure. In this study, by silencing endogenous PLD isozymes using small interfering RNA in HeLa cells, we demonstrate that endogenous PLD1 is required for the normal organization of the actin cytoskeleton, and, more importantly, for cell motility. PLD1 silencing in HeLa cells resulted in dramatic changes in cellular morphology, including the accumulation of stress fibers, as well as cell elongation and flattening, which appeared to be caused by an increased number of focal adhesions, which ultimately culminated in enhanced cell-substratum interactions. Accordingly, serum-induced cell migration was profoundly inhibited by PLD1-silencing. Moreover, the augmented cell substratum interaction and retarded cell migration induced by PLD1-silencing could be restored by the adding back not only of wild type, but also of lipase-inactive PLD1 into knockdown cells. Taken together, our results strongly suggest that endogenous PLD1 is a critical factor in the organization of the actin-based cytoskeleton, with regard to cell adhesion and migration. These effects of PLD1 appear to operate in a lipase activity-independent manner. We also discuss the regulation of Src family kinases by PLD1, as related to the modulation of Pyk2 and cell migration.
Kim
Jung Hwan
JH
Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang, 790-784, South Korea.
Kim
Hyun-wook
HW
Jeon
Hyeona
H
Suh
Pann-Ghill
PG
Ryu
Sung Ho
SH
D001483:Base Sequence; D002465:Cell Movement / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D008049:Lipase / Q000378:metabolism*; D009841:Oligonucleotides; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D034622:RNA Interference
D009841:Oligonucleotides; D008049:Lipase; D010739:Phospholipase D; C106801:phospholipase D1