The Drosophila protein palmitoylome: Characterizing palmitoyl-thioesterases and DHHC palmitoyl-transferases

Palmitoylation is the post-translational addition of a palmitate moiety to a cysteine residue through a covalent thioester bond. The addition and removal of this modification is controlled by both palmitoyl acyl-transferases and thioesterases. Using bioinformatic analysis, we identified 22 DHHC fami...

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Published in:	Fly (Austin, Tex.) Vol. 2; no. 4; pp. 198 - 214
Main Authors:	Bannan, Barbra A., Van Etten, Jamie, Kohler, John A., Tsoi, Yui, Hansen, Nicole M., Sigmon, Stacey, Fowler, Elizabeth, Buff, Haley, Williams, Tiffany S., Ault, Jeffrey G., Glaser, Robert L., Korey, Christopher A.
Format:	Journal Article
Language:	English
Published:	United States Taylor & Francis 01-07-2008
Subjects:	Acyltransferases - genetics Acyltransferases - metabolism Amino Acid Motifs Amino Acid Sequence Animals Binding Biology Bioscience Calcium Cancer Cell Cell Line Cycle Drosophila - enzymology Drosophila - genetics Drosophila - growth & development Drosophila Proteins - genetics Drosophila Proteins - metabolism Genes, Insect Humans Landes Male Membrane Proteins - genetics Membrane Proteins - metabolism Molecular Sequence Data Multigene Family Organogenesis Palmitoyl-CoA Hydrolase - genetics Palmitoyl-CoA Hydrolase - metabolism Proteins Thiolester Hydrolases - genetics Thiolester Hydrolases - metabolism
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Summary:	Palmitoylation is the post-translational addition of a palmitate moiety to a cysteine residue through a covalent thioester bond. The addition and removal of this modification is controlled by both palmitoyl acyl-transferases and thioesterases. Using bioinformatic analysis, we identified 22 DHHC family palmitoyl acyl-transferase homologs in the Drosophila genome. We used in situ hybridization, RT-PCR, and published FlyAtlas microarray data to characterize the expression patterns of all 22 fly homologs. Our results indicate that all are expressed genes, but several, including CG1407, CG4676, CG5620, CG6017/dHIP14, CG6618, CG6627, and CG17257 appear to be enriched in neural tissues suggesting that they are important for neural function. Furthermore, we have found that several may be expressed in a sex-specific manner with adult male-specific expression of CG4483 and CG17195. Using tagged versions of the DHHC genes, we demonstrate that fly DHHC proteins are primarily located in either the Golgi Apparatus or Endoplasmic Reticulum in S2 cells, except for CG1407, which was found on the plasma membrane. We also characterized the subcellular localization and expression of the three known thioesterases: Palmitoyl-protein Thioesterase 1 (Ppt1), Palmitoyl-protein Thioesterase 2 (Ppt2), and Acyl-protein Thioesterase 1 (APT1). Our results indicate that Ppt1 and Ppt2 are the major lysosomal thioesterases while APT1 is the likely cytoplasmic thioesterase. Finally, in vivo rescue experiments show that Ppt2 expression cannot rescue the neural inclusion phenotypes associated with loss of Ppt1, further supporting distinct functions and substrates for these two thioesterases. These results will serve as the basis for a more complete understanding of the protein palmitoylome's normal cellular functions in the fly and will lead to further insights into the molecular etiology of diseases associated with the mis-regulation of palmitoylation.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 These individuals contributed equally to this work
ISSN:	1933-6934 1933-6942
DOI:	10.4161/fly.6621