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Cloning and direct G-protein regulation of phospholipase D from tobacco

Phospholipase D (PLD) and heterotrimeric G-proteins are involved in plant signal transduction pathways at the plasma membrane. There is evidence suggesting that PLD acts downstream from G-proteins, but a direct interaction of specific members has not been shown. In the present paper, a PLD cDNA clon...

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Bibliographic Details
Published in:Biochimica et biophysica acta 2001-02, Vol.1530 (2), p.172-183
Main Authors: Lein, Wolfgang, Saalbach, Gerhard
Format: Article
Language:English
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Summary:Phospholipase D (PLD) and heterotrimeric G-proteins are involved in plant signal transduction pathways at the plasma membrane. There is evidence suggesting that PLD acts downstream from G-proteins, but a direct interaction of specific members has not been shown. In the present paper, a PLD cDNA clone was isolated from tobacco, expressed as a GST fusion in bacteria, and the recombinant protein was purified by glutathione affinity. Its enzymatic properties identified it as an α-type PLD. The α-subunit of a G-protein from tobacco was isolated in a similar way. Both proteins were functional in biochemical assays. When the G-protein was included in the PLD assay, a strong dosage-dependent inhibition of the PLD activity was observed. Different control proteins did not exhibit this inhibitory effect. When GST-NtGPα1 was activated by incubation with GTPγS the inhibitory activity was greatly reduced. These results provide a first indication for a direct regulation of PLDα by a heterotrimeric G-protein α-subunit in plants.
ISSN:1388-1981
0006-3002
1879-2618
DOI:10.1016/S1388-1981(00)00182-7