A novel arachidonic acid-related thioesterase involved in acute steroidogenesis.
We have reported the purification of a phosphoprotein (p43) intermediary in arachidonic acid release and steroid synthesis. Here we describe the cloning and sequencing of a cDNA encoding p43 as well as the hormonal regulation of the p43 transcript. The protein is homologous to a family of novel acyl-CoA thioesterases and identical to a peroxisome proliferator-inducible mitochondrial acyl-CoA thioesterase that shows highest substrate specificity for very-long-chain fatty acids such as arachidoyl- and palmitoyl-CoA. The deduced amino acid sequence of the protein has consensus sites for phosphorylation by different protein kinases, and a putative lipase serine motif. This motif is conserved in several species such as mouse, rat and human. Antibodies raised against a synthetic peptide that includes the lipase serine motif block the action of the protein. The transcript was induced by in vivo stimulation of the adrenals with ACTH. The effect of ACTH was rapid (5 min), reached a maximum (62%) at 15 min and returned to basal levels at 30 min. The effect was inhibited by actinomycin D and enhanced by cycloheximide. Our results provide the first evidence linking acyl-CoA thioesterases, with specificity for very-long-chain acids, and a protein intermediary in steroid synthesis, thereby supporting a regulatory role for acyl-CoA thioesterases in steroidogenic tissues. Given the obligatory role of the protein in the activation of steroidogenesis through arachidonic acid release, we propose the name Arachidonic acid- Related Thioesterase Involved in Steroidogenesis (ARTISt) for p43.
Finkielstein
C V
CV
Department of Biochemistry, School of Medicine, University of Buenos Aires, Argentina.
Maloberti
P
P
Mendez
C F
CF
Podestá
E J
EJ
D000311:Adrenal Glands / Q000187:drug effects; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D016384:Consensus Sequence / Q000235:genetics; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D003609:Dactinomycin / Q000494:pharmacology; D004357:Drug Synergism; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010170:Palmitoyl-CoA Hydrolase; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013256:Steroids / Q000096:biosynthesis*; D013869:Thiolester Hydrolases / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors
D018076:DNA, Complementary; D024101:Mitochondrial Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D012333:RNA, Messenger; D013256:Steroids; D003609:Dactinomycin; D016718:Arachidonic Acid; D000324:Adrenocorticotropic Hormone; D003513:Cycloheximide; D013869:Thiolester Hydrolases; C546434:Acot2 protein, rat; D010170:Palmitoyl-CoA Hydrolase