Differential regulation of Saccharomyces cerevisiae phospholipase D in sporulation and Sec14-independent secretion.
Saccharomyces cerevisiae Spo14, a phosphatidylcholine-specific, phosphatidylinositol (4,5) bisphosphate-activated phospholipase D (PLD), is essential for meiosis and spore formation. Spo14 is also required for secretion in the absence of the phosphatidylinositol/phosphatidylcholine transfer protein Sec14 (i.e., Sec14-independent secretion). In sporulating cells Spo14 is phosphorylated and relocalized within the cell. In contrast, Spo14 does not relocalize and is not phosphorylated in Sec14-independent secretion. Analysis of a partially phosphatidylinositol (4,5) bisphosphate-activated Spo14 mutant, spo14(R894G), revealed that Spo14 function in Sec14-independent secretion, unlike the situation in meiosis, requires fully stimulated PLD activity. Consistent with the differential regulation of Spo14 function during sporulation and secretion, we isolated a mutant allele, spo14-S251P, the product of which is improperly phosphorylated and fails to relocalize and rescue the sporulation phenotype of homozygous spo14 diploids, but supports Sec14-independent secretion. Furthermore, we show that the N-terminal domain of Spo14 is both phosphorylated and sufficient for prospore membrane localization during sporulation. These data indicate that Spo14 phosphorylation and relocalization are essential for the process of sporulation, but dispensable for Sec14-independent secretion. Finally, we demonstrate that Spo14 phosphorylation and relocalization are initiated by nitrogen and glucose limitation and occur independently of the process of meiosis.
Rudge
Simon A
SA
Department of Pharmacological Sciences, State University of New York, Stony Brook, New York 11794-8651, USA.
Zhou
Chun
C
Engebrecht
JoAnne
J
D002352:Carrier Proteins / Q000502:physiology; D004789:Enzyme Activation; D005947:Glucose / Q000378:metabolism; D006358:Hot Temperature; D008565:Membrane Proteins; D009584:Nitrogen / Q000378:metabolism; D010739:Phospholipase D / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D010766:Phosphorylation; D010770:Phosphotransferases; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins; D013172:Spores, Fungal / Q000502:physiology*
D002352:Carrier Proteins; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; C483438:SEC14 protein, S cerevisiae; C118289:SEC24 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D010770:Phosphotransferases; D010739:Phospholipase D; D005947:Glucose; D009584:Nitrogen