SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae

Simon A. Rudge, Trevor R. Pettitt, Chun Zhou, Michael J O Wakelam, JoAnne Engebrecht

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

In Saccharomyces cerevisiae, phospholipase D (PLD), encoded by the SPO14 gene, catalyzes the hydrolysis of phosphatidylcholine, producing choline and phosphatidic acid. SPO14 is essential for cellular differentiation during meiosis and is required for Golgi function when the normal secretory apparatus is perturbed (Sec14-independent secretion). We isolated specific alleles of SPO14 that support Sec14-independent secretion but not sporulation. Identification of these separation-of-function alleles indicates that the role of PLD in these two physiological processes is distinct. Analyses of the mutants reveal that the corresponding proteins are stable, phosphorylated, catalytically active in vitro, and can localize properly within the cell during meiosis. Surprisingly, the separation-of-function mutations map to the conserved catalytic region of the PLD protein. Choline and phosphatidic acid molecular species profiles during Sec14-independent secretion and meiosis reveal that while strains harboring one of these alleles, spo14S-11, hydrolyze phosphatidylcholine in Sec14-independent secretion, they fail to do so during sporulation or normal vegetative growth. These results demonstrate that Spo14 PLD catalytic activity and cellular function can be differentially regulated at the level of phosphatidylcholine hydrolysis.

Original languageEnglish (US)
Pages (from-to)1431-1444
Number of pages14
JournalGenetics
Volume158
Issue number4
StatePublished - 2001
Externally publishedYes

Fingerprint

Phospholipase D
Saccharomyces cerevisiae
Meiosis
Phosphatidylcholines
Phosphatidic Acids
Mutation
Alleles
Choline
Hydrolysis
Physiological Phenomena
Catalytic Domain
Proteins
Growth
Genes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Rudge, S. A., Pettitt, T. R., Zhou, C., Wakelam, M. J. O., & Engebrecht, J. (2001). SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae. Genetics, 158(4), 1431-1444.

SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae. / Rudge, Simon A.; Pettitt, Trevor R.; Zhou, Chun; Wakelam, Michael J O; Engebrecht, JoAnne.

In: Genetics, Vol. 158, No. 4, 2001, p. 1431-1444.

Research output: Contribution to journalArticle

Rudge, SA, Pettitt, TR, Zhou, C, Wakelam, MJO & Engebrecht, J 2001, 'SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae', Genetics, vol. 158, no. 4, pp. 1431-1444.
Rudge, Simon A. ; Pettitt, Trevor R. ; Zhou, Chun ; Wakelam, Michael J O ; Engebrecht, JoAnne. / SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae. In: Genetics. 2001 ; Vol. 158, No. 4. pp. 1431-1444.
@article{8d0d95d130874c50876e8f886bb9846a,
title = "SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae",
abstract = "In Saccharomyces cerevisiae, phospholipase D (PLD), encoded by the SPO14 gene, catalyzes the hydrolysis of phosphatidylcholine, producing choline and phosphatidic acid. SPO14 is essential for cellular differentiation during meiosis and is required for Golgi function when the normal secretory apparatus is perturbed (Sec14-independent secretion). We isolated specific alleles of SPO14 that support Sec14-independent secretion but not sporulation. Identification of these separation-of-function alleles indicates that the role of PLD in these two physiological processes is distinct. Analyses of the mutants reveal that the corresponding proteins are stable, phosphorylated, catalytically active in vitro, and can localize properly within the cell during meiosis. Surprisingly, the separation-of-function mutations map to the conserved catalytic region of the PLD protein. Choline and phosphatidic acid molecular species profiles during Sec14-independent secretion and meiosis reveal that while strains harboring one of these alleles, spo14S-11, hydrolyze phosphatidylcholine in Sec14-independent secretion, they fail to do so during sporulation or normal vegetative growth. These results demonstrate that Spo14 PLD catalytic activity and cellular function can be differentially regulated at the level of phosphatidylcholine hydrolysis.",
author = "Rudge, {Simon A.} and Pettitt, {Trevor R.} and Chun Zhou and Wakelam, {Michael J O} and JoAnne Engebrecht",
year = "2001",
language = "English (US)",
volume = "158",
pages = "1431--1444",
journal = "Genetics",
issn = "0016-6731",
publisher = "Genetics Society of America",
number = "4",

}

TY - JOUR

T1 - SPO14 separation-of-function mutations define unique roles for phospholipase D in secretion and cellular differentiation in Saccharomyces cerevisiae

AU - Rudge, Simon A.

AU - Pettitt, Trevor R.

AU - Zhou, Chun

AU - Wakelam, Michael J O

AU - Engebrecht, JoAnne

PY - 2001

Y1 - 2001

N2 - In Saccharomyces cerevisiae, phospholipase D (PLD), encoded by the SPO14 gene, catalyzes the hydrolysis of phosphatidylcholine, producing choline and phosphatidic acid. SPO14 is essential for cellular differentiation during meiosis and is required for Golgi function when the normal secretory apparatus is perturbed (Sec14-independent secretion). We isolated specific alleles of SPO14 that support Sec14-independent secretion but not sporulation. Identification of these separation-of-function alleles indicates that the role of PLD in these two physiological processes is distinct. Analyses of the mutants reveal that the corresponding proteins are stable, phosphorylated, catalytically active in vitro, and can localize properly within the cell during meiosis. Surprisingly, the separation-of-function mutations map to the conserved catalytic region of the PLD protein. Choline and phosphatidic acid molecular species profiles during Sec14-independent secretion and meiosis reveal that while strains harboring one of these alleles, spo14S-11, hydrolyze phosphatidylcholine in Sec14-independent secretion, they fail to do so during sporulation or normal vegetative growth. These results demonstrate that Spo14 PLD catalytic activity and cellular function can be differentially regulated at the level of phosphatidylcholine hydrolysis.

AB - In Saccharomyces cerevisiae, phospholipase D (PLD), encoded by the SPO14 gene, catalyzes the hydrolysis of phosphatidylcholine, producing choline and phosphatidic acid. SPO14 is essential for cellular differentiation during meiosis and is required for Golgi function when the normal secretory apparatus is perturbed (Sec14-independent secretion). We isolated specific alleles of SPO14 that support Sec14-independent secretion but not sporulation. Identification of these separation-of-function alleles indicates that the role of PLD in these two physiological processes is distinct. Analyses of the mutants reveal that the corresponding proteins are stable, phosphorylated, catalytically active in vitro, and can localize properly within the cell during meiosis. Surprisingly, the separation-of-function mutations map to the conserved catalytic region of the PLD protein. Choline and phosphatidic acid molecular species profiles during Sec14-independent secretion and meiosis reveal that while strains harboring one of these alleles, spo14S-11, hydrolyze phosphatidylcholine in Sec14-independent secretion, they fail to do so during sporulation or normal vegetative growth. These results demonstrate that Spo14 PLD catalytic activity and cellular function can be differentially regulated at the level of phosphatidylcholine hydrolysis.

UR - http://www.scopus.com/inward/record.url?scp=0034850605&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034850605&partnerID=8YFLogxK

M3 - Article

C2 - 11514437

AN - SCOPUS:0034850605

VL - 158

SP - 1431

EP - 1444

JO - Genetics

JF - Genetics

SN - 0016-6731

IS - 4

ER -