The contribution of serine 194 phosphorylation to steroidogenic acute regulatory protein function.
The steroidogenic acute regulatory protein (StAR) facilitates the delivery of cholesterol to the inner mitochondrial membrane, where the cholesterol side-chain cleavage enzyme catalyzes the initial step of steroid hormone biosynthesis. StAR was initially identified in adrenocortical cells as a phosphoprotein, the expression and phosphorylation of which were stimulated by corticotropin. A number of in vitro studies have implicated cAMP-dependent phosphorylation at serine 194 (S194, S195 in human StAR) as an important residue for StAR activity. To explore the importance of S194 phosphorylation in StAR function in vivo, we developed a transgenic model using a bacterial artificial chromosome expressing either wild-type (WT) StAR or StAR mutation S194A to rescue StAR knockout (KO) mice. Despite StAR protein expression comparable to or higher than amounts seen with control animals or rescue with WT StAR, S194A StAR did not rescue the neonatal lethality and only partially rescued the sex reversal in male mice observed uniformly in StAR KO mice. Like the StAR KO mice, the adrenal cortex and testicular Leydig cells contained abundant lipid deposits when stained with oil red O. Adrenal StAR from S194A rescue animals lacks an acidic species, which appears upon corticotropin stimulation in animals rescued with WT StAR, consistent with defective StAR phosphorylation. These findings demonstrate that S194 is an essential residue for normal StAR function in the adrenal cortex and testes of mice.
Department of Pediatrics (G.S.), Tokyo Dental College Ichikawa General Hospital, Ichikawa, Chiba, 272-8513, Japan; Department of Internal Medicine (G.S., M.Z., R.J.A.), University of Texas Southwestern Medical Center, Dallas, Texas 75390; Department of Pediatrics (T.I., T.M., T.H.), Keio University School of Medicine, Tokyo, 160-8582, Japan; PAPRSB Institute of Health Science (M.Z.), University of Brunei Darussalam, Brunei; and Department of Internal Medicine (R.J.A.), University of Michigan, Ann Arbor, Michigan 48109.
Goro
G
Sasaki
Mohamad
M
Zubair
Tomohiro
T
Ishii
Toshikatsu
T
Mitsui
Tomonobu
T
Hasegawa
Richard J
RJ
Auchus
D000302:Adrenal Cortex / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000378:metabolism; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D002784:Cholesterol / Q000378:metabolism; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D005260:Female; D012739:Gonadal Steroid Hormones / Q000096:biosynthesis*; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D051336:Mitochondrial Membranes / Q000201:enzymology / Q000378:metabolism; D009154:Mutation; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D000098682:Steroidogenic Acute Regulatory Protein
D012739:Gonadal Steroid Hormones; D010750:Phosphoproteins; D000098682:Steroidogenic Acute Regulatory Protein; D000324:Adrenocorticotropic Hormone; D002784:Cholesterol; D002786:Cholesterol Side-Chain Cleavage Enzyme