Cytosolic and mitochondrial proteins as possible targets of cycloheximide effect on adrenal steroidogenesis.
It is well accepted that protein(s) with a short half-life are required in the pathway leading to steroid synthesis following stimulation by trophic hormones. A correlation between the disappearance of several proteins in different subcellular compartments and the inhibition of steroid synthesis produced by cycloheximide (CHx) has also been shown. In the present report we describe the effect of CHx in the stimulation of steroid synthesis using a cell-free assay. Mitochondrial progesterone (P4) production was studied by recombination of the different subcellular fractions of adrenal zona fasciculata and determined by radioimmunoassay. Soluble factors from ACTH-treated adrenals produced a four-fold stimulation of mitochondrial steroidogenesis (3.0 +/- 0.6 vs. 13.3 +/- 0.5 ng P4/tube for control and ACTH-treated adrenals respectively). Mitochondria obtained from CHx-ACTH-treated adrenals fail to respond to soluble ACTH-dependent factors. A permeable analogue of cholesterol (22(R)-OH cholesterol) could overcome the inhibition imposed by CHx, confirming the role of mitochondrial proteins in intramitochondrial cholesterol transport. The treatment of the adrenals with CHx 10 minutes before ACTH administration abolished also the stimulation induced by the cytosol on control mitochondria (2.6 +/- 0.5 vs. 13.0 +/- 1.0 ng P4/tube for CHx-ACTH-treated cytosol vs. ACTH-treated cytosol). Arachidonic acid (AA) added to CHx-ACTH-treated cytosol subdued this inhibition (10.3 +/- 1.2 ng P4/tube). CHx treatment had no effect on the stimulation by ACTH of the cAMP-dependent protein kinase. These results indicate the involvement of a cycloheximide-sensitive protein in the release of AA in adrenal steroidogenesis.
Dada
L
L
Department of Biochemistry, School of Medicine, University of Buenos Aires, Argentina.
Cornejo Maciel
F
F
Neuman
I
I
Mele
P G
PG
Maloberti
P
P
Paz
C
C
Cymeryng
C
C
Finkielstein
C
C
Mendez
C F
CF
Podestá
E J
EJ
D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D011374:Progesterone / Q000096:biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013256:Steroids / Q000096:biosynthesis*; D015383:Zona Fasciculata / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*
D011500:Protein Synthesis Inhibitors; D011506:Proteins; D013256:Steroids; D016718:Arachidonic Acid; D011374:Progesterone; D003513:Cycloheximide; D002786:Cholesterol Side-Chain Cleavage Enzyme