JCB logo
Immu
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 544K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Rudge, S. A.
Right arrow Articles by Engebrecht, J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rudge, S. A.
Right arrow Articles by Engebrecht, J.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/1998//81 $5.00
The Journal of Cell Biology, Volume 140, Number 1, , 1998 81-90

Article

Relocalization of Phospholipase D Activity Mediates Membrane Formation During Meiosis



Simon A. Rudge, Andrew J. Morris, and JoAnne Engebrecht

Department of Pharmacological Sciences, State University of New York, Stony Brook, Stony Brook, New York 11794-8651

Phospholipase D (PLD) enzymes catalyze the hydrolysis of phosphatidylcholine and are involved in membrane trafficking and cytoskeletal reorganization. The Saccharomyces cerevisiae SPO14 gene encodes a PLD that is essential for meiosis. We have analyzed the role of PLD in meiosis by examining two mutant proteins, one with a point mutation in a conserved residue (Spo14pK-> H) and one with an amino-terminal deletion (Spo14p{Delta}N), neither of which can restore meiosis in a spo14 deletion strain. Spo14pK-> H is enzymatically inactive, indicating that PLD activity is required, whereas Spo14p{Delta}N retains PLD catalytic activity in vitro, indicating that PLD activity is not sufficient for meiosis. To explore other aspects of Spo14 function, we followed the localization of the enzyme during meiosis. Spo14p is initially distributed throughout the cell, becomes concentrated at the spindle pole bodies after the meiosis I division, and at meiosis II localizes to the new spore membrane as it surrounds the nuclei and then expands to encapsulate the associated cytoplasm during the formation of spores. The catalytically inactive protein also undergoes relocalization during meiosis; however, in the absence of PLD activity, no membrane is formed. In contrast, Spo14p{Delta}N does not relocalize properly, indicating that the failure of this protein to complement a spo14 mutant is due to its inability to localize its PLD activity. Furthermore, we find that Spo14p movement is correlated with phosphorylation of the protein. These experiments indicate that PLD participates in regulated membrane formation during meiosis, and that both its catalytic activity and subcellular redistribution are essential for this function.

Abbreviations used in this paper: BODIPY-PA, 2-decanoyl-1-(O-[11-{4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3- propionyl}amino] undecyl]-sn-glycero-3-phosphatidate; BODIPY-PC, 2-decanoyl-1-(O-[11-{4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3- propionyl}amino] undecyl)-sn-glycero-3-phosphocholine; DAG, diacylglycerol; DAPI, 4'-6'diaminophenylindole; GFP, green fluorescent protein; HA, hemagglutinin; PA, phosphatidic acid; PC, phosphatidylcholine; PIP2, phosphatidylinositol 4,5-bisphosphate; PKC, protein kinase C; PLD, phospholipase D.

Address all correspondence to JoAnne Engebrecht, Department of Pharmacological Sciences, State University of New York, Stony Brook, Stony Brook, NY 11794-8651. Tel.: (516) 444-7815. Fax: (516) 444-3218. E-mail: joanne{at}pharm.sunysb.edu


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents