Inhibition of phospholipase D activity by fodrin. An active role for the cytoskeleton.
Phospholipase D (PLD) is a major enzyme implicated in important cellular processes such as secretion and proliferation. The knowledge of its regulation is essential to understand the control of these phenomena. Several proteins activating PLD have been described in the last years. In this report, we chromatographed bovine brain cytosolic proteins to identify fodrin, the non-erythroid spectrin, as the first described inhibitor of PLD. A cytosolic fraction with an inhibitory effect on PLD activity loses its capacity after immunoprecipitation of fodrin. Moreover, at 1 nM, purified fodrin blocks fully and quickly PLD activity, whatever the stimuli used. In contrast, fodrin has no effect on adenylate cyclase activity. Fodrin-analogous proteins like dimeric or tetrameric erythroid spectrin have the same inhibitory effect on PLD, at higher concentrations. Other cytoskeletal proteins, actin and vimentin, are inefficient on PLD inhibition. The mechanisms implicated in PLD modulation such as post-translational modifications of fodrin and the role of small G-proteins on the cytoskeleton regulation are discussed. In conclusion, this study reveals that fodrin is involved in the control of PLD activity, suggesting that the cytoskeleton could have an active role in control of secretion and proliferation.
Lukowski
S
S
Unité INSERM 332, ICGM, Paris, France.
Lecomte
M C
MC
Mira
J P
JP
Marin
P
P
Gautero
H
H
Russo-Marie
F
F
Geny
B
B
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002154:Calpain / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D003599:Cytoskeleton / Q000201:enzymology*; D004789:Enzyme Activation; D006160:Guanosine Triphosphate / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D010739:Phospholipase D / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000378:metabolism; D015398:Signal Transduction; D013049:Spectrin / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism
D000262:Adenylyl Cyclases; D002118:Calcium; D002147:Calmodulin; D002154:Calpain; D002352:Carrier Proteins; D006160:Guanosine Triphosphate; D008840:Microfilament Proteins; D010713:Phosphatidylcholines; D019269:Phosphatidylinositol 4,5-Diphosphate; D010739:Phospholipase D; D010741:Phospholipases A; D013049:Spectrin; D013755:Tetradecanoylphorbol Acetate; D010738:Type C Phospholipases