HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Arakane, F. Articles by Strauss III, J. F. Search for Related Content PubMed PubMed Citation Articles by Arakane, F. Articles by Strauss III, J. F. Social Bookmarking                      What's this?

J Biol Chem, Vol. 273, Issue 26, 16339-16345, June 26, 1998

The Mechanism of Action of Steroidogenic Acute Regulatory Protein (StAR)
StAR ACTS ON THE OUTSIDE OF MITOCHONDRIA TO STIMULATE STEROIDOGENESIS

Futoshi Arakane, Caleb B. Kallen, Hidemichi Watari, James A. Foster, Naresh Babu V. Sepuri^§, Debkumar Pain^§, Steven E. Stayrook^¶, Mitchell Lewis^¶, George L. Gerton, and Jerome F. Strauss III^§

From the  Center for Research on Reproduction and Women's Health and the Department of Obstetrics and Gynecology, ^§ Physiology, and ^¶ Biochemistry and Biophysics, University of Pennsylvania Medical Center, Philadelphia, Pennsylvania 19104

Steroidogenic acute regulatory protein (StAR) plays an essential role in steroidogenesis, facilitating delivery of cholesterol to cytochrome P450_scc on the inner mitochondrial membrane. StAR is synthesized in the cytoplasm and is subsequently imported by mitochondria and processed to a mature form by cleavage of the NH₂-terminal mitochondrial targeting sequence. To explore the mechanism of StAR action, we produced 6-histidine-tagged N-62 StAR (His-tag StAR) constructs lacking the NH₂-terminal 62 amino acids that encode the mitochondrial targeting sequence and examined their steroidogenic activity in intact cells and on isolated mitochondria. His-tag StAR proteins stimulated pregnenolone synthesis to the same extent as wild-type StAR when expressed in COS-1 cells transfected with the cholesterol side-chain cleavage system. His-tag StAR was diffusely distributed in the cytoplasm of transfected COS-1 cells whereas wild-type StAR was localized to mitochondria. There was no evidence at the light or electron microscope levels for selective localization of His-tag StAR protein to mitochondrial membranes. In vitro import assays demonstrated that wild-type StAR preprotein was imported and processed to mature protein that was protected from subsequent trypsin treatment. In contrast, His-tag StAR was not imported and protein associated with mitochondria was sensitive to trypsin. Using metabolically labeled COS-1 cells transfected with wild-type or His-tag StAR constructs, we confirmed that wild-type StAR preprotein was imported and processed by mitochondria, whereas His-tag StAR remained largely cytosolic and unprocessed. To determine whether cytosolic factors are required for StAR action, we developed an assay system using washed mitochondria isolated from bovine corpora lutea and purified recombinant His-tag StAR proteins expressed in Escherichia coli. Recombinant His-tag StAR stimulated pregnenolone production in a dose- and time-dependent manner, functioning at nanomolar concentrations. A point mutant of StAR (A218V) that causes lipoid congenital adrenal hyperplasia was incorporated into the His-tag protein. This mutant was steroidogenically inactive in COS-1 cells and on isolated mitochondria. Our observations conclusively document that StAR acts on the outside of mitochondria, independent of mitochondrial import, and in the absence of cytosol. The ability to produce bioactive recombinant StAR protein paves the way for refined structural studies of StAR and StAR mutants.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				P. R. Manna, M. T. Dyson, and D. M. Stocco Regulation of the steroidogenic acute regulatory protein gene expression: present and future perspectives Mol. Hum. Reprod., June 1, 2009; 15(6): 321 - 333. [Abstract] [Full Text] [PDF]

				J. A. Locke, E. S. Guns, A. A. Lubik, H. H. Adomat, S. C. Hendy, C. A. Wood, S. L. Ettinger, M. E. Gleave, and C. C. Nelson Androgen Levels Increase by Intratumoral De novo Steroidogenesis during Progression of Castration-Resistant Prostate Cancer Cancer Res., August 1, 2008; 68(15): 6407 - 6415. [Abstract] [Full Text] [PDF]

				G. Sasaki, T. Ishii, P. Jeyasuria, Y. Jo, A. Bahat, J. Orly, T. Hasegawa, and K. L. Parker Complex Role of the Mitochondrial Targeting Signal in the Function of Steroidogenic Acute Regulatory Protein Revealed by Bacterial Artificial Chromosome Transgenesis in Vivo Mol. Endocrinol., April 1, 2008; 22(4): 951 - 964. [Abstract] [Full Text] [PDF]

				M. T Dyson, J. K Jones, M. P Kowalewski, P. R Manna, M. Alonso, M. E Gottesman, and D. M Stocco Mitochondrial A-Kinase Anchoring Protein 121 Binds Type II Protein Kinase A and Enhances Steroidogenic Acute Regulatory Protein-Mediated Steroidogenesis in MA-10 Mouse Leydig Tumor Cells Biol Reprod, February 1, 2008; 78(2): 267 - 277. [Abstract] [Full Text] [PDF]

				Z. Granot, O. Kobiler, N. Melamed-Book, S. Eimerl, A. Bahat, B. Lu, S. Braun, M. R. Maurizi, C. K. Suzuki, A. B. Oppenheim, et al. Turnover of Mitochondrial Steroidogenic Acute Regulatory (StAR) Protein by Lon Protease: The Unexpected Effect of Proteasome Inhibitors Mol. Endocrinol., September 1, 2007; 21(9): 2164 - 2177. [Abstract] [Full Text] [PDF]

				A Stigliano, O Gandini, L Cerquetti, P Gazzaniga, S Misiti, S Monti, A Gradilone, P Falasca, M Poggi, E Brunetti, et al. Increased metastatic lymph node 64 and CYP17 expression are associated with high stage prostate cancer J. Endocrinol., July 1, 2007; 194(1): 55 - 61. [Abstract] [Full Text] [PDF]

				W. L. Miller StAR Search--What We Know about How the Steroidogenic Acute Regulatory Protein Mediates Mitochondrial Cholesterol Import Mol. Endocrinol., March 1, 2007; 21(3): 589 - 601. [Abstract] [Full Text] [PDF]

				M. Murcia, J. D. Faraldo-Gomez, F. R. Maxfield, and B. Roux Modeling the structure of the StART domains of MLN64 and StAR proteins in complex with cholesterol J. Lipid Res., December 1, 2006; 47(12): 2614 - 2630. [Abstract] [Full Text] [PDF]

				P. R. Manna, S. P. Chandrala, S. R. King, Y. Jo, R. Counis, I. T. Huhtaniemi, and D. M. Stocco Molecular Mechanisms of Insulin-like Growth Factor-I Mediated Regulation of the Steroidogenic Acute Regulatory Protein in Mouse Leydig Cells Mol. Endocrinol., February 1, 2006; 20(2): 362 - 378. [Abstract] [Full Text] [PDF]

				B. Y. Baker, D. C. Yaworsky, and W. L. Miller A pH-dependent Molten Globule Transition Is Required for Activity of the Steroidogenic Acute Regulatory Protein, StAR J. Biol. Chem., December 16, 2005; 280(50): 41753 - 41760. [Abstract] [Full Text] [PDF]

				R. E. Soccio, R. M. Adams, K. N. Maxwell, and J. L. Breslow Differential Gene Regulation of StarD4 and StarD5 Cholesterol Transfer Proteins: ACTIVATION OF StarD4 BY STEROL REGULATORY ELEMENT-BINDING PROTEIN-2 AND StarD5 BY ENDOPLASMIC RETICULUM STRESS J. Biol. Chem., May 13, 2005; 280(19): 19410 - 19418. [Abstract] [Full Text] [PDF]

				F. Alpy, V. K. Latchumanan, V. Kedinger, A. Janoshazi, C. Thiele, C. Wendling, M.-C. Rio, and C. Tomasetto Functional Characterization of the MENTAL Domain J. Biol. Chem., May 6, 2005; 280(18): 17945 - 17952. [Abstract] [Full Text] [PDF]

				D. C. Yaworsky, B. Y. Baker, H. S. Bose, K. B. Best, L. B. Jensen, J. D. Bell, M. A. Baldwin, and W. L. Miller pH-dependent Interactions of the Carboxyl-terminal Helix of Steroidogenic Acute Regulatory Protein with Synthetic Membranes J. Biol. Chem., January 21, 2005; 280(3): 2045 - 2054. [Abstract] [Full Text] [PDF]

				T. Kishida, I. Kostetskii, Z. Zhang, F. Martinez, P. Liu, S. U. Walkley, N. K. Dwyer, E. J. Blanchette-Mackie, G. L. Radice, and J. F. Strauss III Targeted Mutation of the MLN64 START Domain Causes Only Modest Alterations in Cellular Sterol Metabolism J. Biol. Chem., April 30, 2004; 279(18): 19276 - 19285. [Abstract] [Full Text] [PDF]

				Z. Granot, R. Geiss-Friedlander, N. Melamed-Book, S. Eimerl, R. Timberg, A. M. Weiss, K. H. Hales, D. B. Hales, D. M. Stocco, and J. Orly Proteolysis of Normal and Mutated Steroidogenic Acute Regulatory Proteins in the Mitochondria: the Fate of Unwanted Proteins Mol. Endocrinol., December 1, 2003; 17(12): 2461 - 2476. [Abstract] [Full Text] [PDF]

				T. Sugawara, H. Shimizu, N. Hoshi, A. Nakajima, and S. Fujimoto Steroidogenic Acute Regulatory Protein-binding Protein Cloned by a Yeast Two-hybrid System J. Biol. Chem., October 24, 2003; 278(43): 42487 - 42494. [Abstract] [Full Text] [PDF]

				H. S. Sun, K.-Y. Hsiao, C.-C. Hsu, M.-H. Wu, and S.-J. Tsai Transactivation of Steroidogenic Acute Regulatory Protein in Human Endometriotic Stromal Cells Is Mediated by the Prostaglandin EP2 Receptor Endocrinology, September 1, 2003; 144(9): 3934 - 3942. [Abstract] [Full Text] [PDF]

				T. Diemer, J. A. Allen, K. H. Hales, and D. B. Hales Reactive Oxygen Disrupts Mitochondria in MA-10 Tumor Leydig Cells and Inhibits Steroidogenic Acute Regulatory (StAR) Protein and Steroidogenesis Endocrinology, July 1, 2003; 144(7): 2882 - 2891. [Abstract] [Full Text] [PDF]

				K. Tajima, A. Dantes, Z. Yao, K. Sorokina, F. Kotsuji, R. Seger, and A. Amsterdam Down-Regulation of Steroidogenic Response to Gonadotropins in Human and Rat Preovulatory Granulosa Cells Involves Mitogen-Activated Protein Kinase Activation and Modulation of DAX-1 and Steroidogenic Factor-1 J. Clin. Endocrinol. Metab., May 1, 2003; 88(5): 2288 - 2299. [Abstract] [Full Text] [PDF]

				R. C. Tuckey, M. J. Headlam, H. S. Bose, and W. L. Miller Transfer of Cholesterol between Phospholipid Vesicles Mediated by the Steroidogenic Acute Regulatory Protein (StAR) J. Biol. Chem., November 27, 2002; 277(49): 47123 - 47128. [Abstract] [Full Text] [PDF]

				H. Takemori, Y. Katoh, N. Horike, J. Doi, and M. Okamoto ACTH-induced Nucleocytoplasmic Translocation of Salt-inducible Kinase. IMPLICATION IN THE PROTEIN KINASE A-ACTIVATED GENE TRANSCRIPTION IN MOUSE ADRENOCORTICAL TUMOR CELLS J. Biol. Chem., October 25, 2002; 277(44): 42334 - 42343. [Abstract] [Full Text] [PDF]

				K. Tajima, K. Hosokawa, Y. Yoshida, A. Dantes, R. Sasson, F. Kotsuji, and A. Amsterdam Establishment of FSH-responsive cell lines by transfection of pre-ovulatory human granulosa cells with mutated p53 (p53val135) and Ha-ras genes Mol. Hum. Reprod., January 1, 2002; 8(1): 48 - 57. [Abstract] [Full Text] [PDF]

				I. P. Artemenko, D. Zhao, D. B. Hales, K. H. Hales, and C. R. Jefcoate Mitochondrial Processing of Newly Synthesized Steroidogenic Acute Regulatory Protein (StAR), but Not Total StAR, Mediates Cholesterol Transfer to Cytochrome P450 Side Chain Cleavage Enzyme in Adrenal Cells J. Biol. Chem., November 30, 2001; 276(49): 46583 - 46596. [Abstract] [Full Text] [PDF]

				F. Schroeder, A. M. Gallegos, B. P. Atshaves, S. M. Storey, A. L. McIntosh, A. D. Petrescu, H. Huang, O. Starodub, H. Chao, H. Yang, et al. Recent Advances in Membrane Microdomains: Rafts, Caveolae, and Intracellular Cholesterol Trafficking Experimental Biology and Medicine, November 1, 2001; 226(10): 873 - 890. [Abstract] [Full Text] [PDF]

				D. M. Stocco Tracking the Role of a StAR in the Sky of the New Millennium Mol. Endocrinol., August 1, 2001; 15(8): 1245 - 1254. [Abstract] [Full Text] [PDF]

				M.-C. Huang and W. L. Miller Creation and Activity of COS-1 Cells Stably Expressing the F2 Fusion of the Human Cholesterol Side-Chain Cleavage Enzyme System Endocrinology, June 1, 2001; 142(6): 2569 - 2576. [Abstract] [Full Text] [PDF]

				P. R. Manna, J. Kero, M. Tena-Sempere, P. Pakarinen, D. M. Stocco, and I. T. Huhtaniemi Assessment of Mechanisms of Thyroid Hormone Action in Mouse Leydig Cells: Regulation of the Steroidogenic Acute Regulatory Protein, Steroidogenesis, and Luteinizing Hormone Receptor Function Endocrinology, January 1, 2001; 142(1): 319 - 331. [Abstract] [Full Text] [PDF]

				N. Sekar, H. A. LaVoie, and J. D. Veldhuis Concerted Regulation of Steroidogenic Acute Regulatory Gene Expression by Luteinizing Hormone and Insulin (or Insulin-Like Growth Factor I) in Primary Cultures of Porcine Granulosa-Luteal Cells Endocrinology, November 1, 2000; 141(11): 3983 - 3992. [Abstract] [Full Text] [PDF]

				P. C. White and P. W. Speiser Congenital Adrenal Hyperplasia due to 21-Hydroxylase Deficiency Endocr. Rev., June 1, 2000; 21(3): 245 - 291. [Abstract] [Full Text]

				J.-G. Lehoux, D. B. Hales, A. Fleury, N. Brière, D. Martel, and L. Ducharme The in Vivo Effects of Adrenocorticotropin and Sodium Restriction on the Formation of Different Species of Steroidogenic Acute Regulatory Protein in Rat Adrenal Endocrinology, November 1, 1999; 140(11): 5154 - 5164. [Abstract] [Full Text]

				H. S. Bose, R. M. Whittal, M. A. Baldwin, and W. L. Miller The active form of the steroidogenic acute regulatory protein, StAR, appears to be a molten globule PNAS, June 22, 1999; 96(13): 7250 - 7255. [Abstract] [Full Text] [PDF]

				A. Kerban, D. Boerboom, and J. Sirois Human Chorionic Gonadotropin Induces an Inverse Regulation of Steroidogenic Acute Regulatory Protein Messenger Ribonucleic Acid in Theca Interna and Granulosa Cells of Equine Preovulatory Follicles Endocrinology, February 1, 1999; 140(2): 667 - 674. [Abstract] [Full Text]

				C. B. Kallen, J. T. Billheimer, S. A. Summers, S. E. Stayrook, M. Lewis, and J. F. Strauss III Steroidogenic Acute Regulatory Protein (StAR) Is A Sterol Transfer Protein J. Biol. Chem., October 9, 1998; 273(41): 26285 - 26288. [Abstract] [Full Text] [PDF]

				F. Alpy, M.-E. Stoeckel, A. Dierich, J.-M. Escola, C. Wendling, M.-P. Chenard, M. T. Vanier, J. Gruenberg, C. Tomasetto, and M.-C. Rio The Steroidogenic Acute Regulatory Protein Homolog MLN64, a Late Endosomal Cholesterol-binding Protein J. Biol. Chem., February 2, 2001; 276(6): 4261 - 4269. [Abstract] [Full Text] [PDF]

				R. Seger, T. Hanoch, R. Rosenberg, A. Dantes, W. E. Merz, J. F. Strauss III, and A. Amsterdam The ERK Signaling Cascade Inhibits Gonadotropin-stimulated Steroidogenesis J. Biol. Chem., April 20, 2001; 276(17): 13957 - 13964. [Abstract] [Full Text] [PDF]

				K. Christensen, H. S. Bose, F. M. Harris, W. L. Miller, and J. D. Bell Binding of Steroidogenic Acute Regulatory Protein to Synthetic Membranes Suggests an Active Molten Globule J. Biol. Chem., May 11, 2001; 276(20): 17044 - 17051. [Abstract] [Full Text] [PDF]

				A. D. Petrescu, A. M. Gallegos, Y. Okamura, J. F. Strauss III, and F. Schroeder Steroidogenic Acute Regulatory Protein Binds Cholesterol and Modulates Mitochondrial Membrane Sterol Domain Dynamics J. Biol. Chem., September 28, 2001; 276(40): 36970 - 36982. [Abstract] [Full Text] [PDF]