In:
Journal of Cell Science, The Company of Biologists, Vol. 119, No. 17 ( 2006-09-01), p. 3613-3621
Abstract:
Phosphatidylinositol-4-kinase-IIIβ (PI4KIIIβ) is activated at the Golgi compartment by PKD-mediated phosphorylation. Subsequent mechanisms responsible for continuous PtdIns(4)P production at Golgi membranes and potential interaction partners of activated PI4KIIIβ are unknown. Here we identify phosphoserine/-threonine binding 14-3-3 proteins as novel regulators of PI4KIIIβ activity downstream of this phosphorylation. The PI4KIIIβ-14-3-3 interaction, evident from GST pulldowns, co-immunoprecipitations and bimolecular fluorescence complementation, was augmented by phosphatase inhibition with okadaic acid. Binding of 14-3-3 proteins to PI4KIIIβ involved the PKD phosphorylation site Ser294, evident from reduced 14-3-3 binding to a S294A PI4KIIIβ mutant. Expression of dominant negative 14-3-3 proteins resulted in decreased PI4KIIIβ Ser294 phosphorylation, whereas wildtype 14-3-3 proteins increased phospho-PI4KIIIβ levels. This was because of protection of PI4KIIIβ Ser294 phosphorylation from phosphatase-mediated dephosphorylation. The functional significance of the PI4KIIIβ-14-3-3 interaction was evident from a reduction of PI4KIIIβ activity upon dominant negative 14-3-3 protein expression. We propose that 14-3-3 proteins function as positive regulators of PI4KIIIβ activity by protecting the lipid kinase from active site dephosphorylation, thereby ensuring a continuous supply of PtdIns(4)P at the Golgi compartment.
Type of Medium:
Online Resource
ISSN:
1477-9137
,
0021-9533
Language:
English
Publisher:
The Company of Biologists
Publication Date:
2006
detail.hit.zdb_id:
219171-4
detail.hit.zdb_id:
1483099-1
SSG:
12
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