Cloning and expression of steroid sulfatase cDNA and the frequent occurrence of deletions in STS deficiency: Implications for X-Y interchange

doi:10.1016/0092-8674(87)90447-8

Cell

Volume 49, Issue 4, 22 May 1987, Pages 443-454

https://doi.org/10.1016/0092-8674(87)90447-8 Get rights and content

Abstract

Human STS is a microsomal enzyme important in steroid metabolism. The gene encoding STS is pseudoautosomal in the mouse but not in humans, and escapes X inactivation in both species. We have prepared monoclonal and polyclonal antibodies to the protein which has been purified and from which partial amino acid sequence data have been obtained. cDNA clones containing the entire coding sequence were isolated, sequenced, and expressed in heterologous cells. Variable length transcripts have been shown to be present and due to usage of alternative poly(A) addition sites. The functional gene maps to Xp22.3-Xpter and there is a pseudogene on Yq suggesting a recent pericentric inversion. Absence of STS enzymatic activity occurs frequently in human populations and produces a visible phenotype of scaly skin or ichthyosis. Ten patients with inherited STS deficiency were studied and eight had complete gene deletions. The possibility that STS deficiency results from aberrant X-Y interchange is discussed.

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Those were indeed halcyon days for gene hunters. It was not until the ESDR was approaching its 18th birthday that the first genetic breakthrough for an inherited skin disease was made: microdeletions in STS were shown to underlie X-linked ichthyosis (Ballabio et al., 1987; Yen et al., 1987). Thereafter, during the 1990s, a plethora of genes for major genodermatoses were identified through genetic linkage studies, including keratin genes KRT5 and KRT14 for epidermolysis bullosa (EB) simplex (Bonifas et al., 1991; Coulombe et al., 1991; Lane et al., 1992) and TGM1 for autosomal recessive congenital ichthyosis (ARCI) (Huber et al., 1995).
Launch of the European Society for Dermatological Research (ESDR) in 1970 coincided with genetics also entering a new era. Arriving alongside new models of DNA structure and the discovery of restriction endonucleases, the ESDR has parallel-tracked 50 years of major developments in genomics, technological innovations, and big data. Patients with rare Mendelian genetic skin diseases have witnessed the discovery of causative genes and pathogenic variants, improved genetic counseling, and the advent of prenatal diagnosis. Translational research has also heralded early phase clinical trials of gene, cell, and protein therapies, as well as enhanced disease models, mechanism-based therapies, and impactful clinical progress.
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We investigated the effects of cord blood PFASs on fetal reproductive hormones and its potential mechanism in relation to steroidogenic enzymes.
Ten selected PFASs (n = 351) including PFOS, PFOA, PFBS, PFDA, PFDoA, PFHpA, PFHxS, PFNA, PFOSA, and PFUA, and two reproductive hormones estradiol (E2) (n = 351) and testosterone (T) (n = 349) were measured in 351 cord blood serum samples from a Chinese birth cohort between 2010 and 2013. Three steroidogenic enzymes including P450arom (n = 125), 3β-HSD1 (n = 123), and 17β-HSD1 (n = 116) were measured in 125 placental tissue samples. Linear regression tested the associations between cord blood PFASs and reproductive hormones in cord blood. Mediation analysis assessed the role of placental steroidogenic enzymes between cord blood PFASs and reproductive hormones.
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Steroid sulfatase (STS) plays an important role in the regulation of steroid hormones. Metabolism of steroid hormones in zebrafish has been investigated, but the action of steroid sulfatase remains unknown. In this study, a zebrafish sts was cloned, expressed, purified, and characterized in comparison with the orthologous human enzyme. Enzymatic assays demonstrated that similar to human STS, zebrafish Sts was most active in catalyzing the hydrolysis of estrone-sulfate and estradiol-sulfate, among five steroid sulfates tested as substrates. Kinetic analyses revealed that the K_m values of zebrafish Sts and human STS differed with respective substrates, but the catalytic efficiency as reflected by the V_max/K_m appeared comparable, except for DHEA-sulfate with which zebrafish Sts appeared less efficient. While zebrafish Sts was catalytically active at 28 °C, the enzyme appeared more active at 37 °C and with similar K_m values to those determined at 28 °C. Assays performed in the presence of different divalent cations showed that the activities of both zebrafish and human STSs were stimulated by Ca²⁺, Mg²⁺, and Mn²⁺, and inhibited by Zn⁺² and Fe²⁺. EMATE and STX64, two known mammalian steroid sulafatase inhibitors, were shown to be capable of inhibiting the activity of zebrafish Sts. Collectively, the results obtained indicated that zebrafish Sts exhibited enzymatic characteristics comparable to the human STS, suggesting that the physiological function of STS may be conserved between zebrafish and humans.
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Dehydroepiandrosterone (DHEA)-SO₄ of adrenal origin is the major C19 steroid in the serum. It is a precursor of intratumoral androgen biosynthesis in patients with advanced prostate cancer following chemical or surgical castration. DHEA is a product of the P450c17 (17α-hydroxylase-17,20-lyase) enzyme. Despite inhibition of P450c17 with new agents, e.g., Abiraterone acetate, Orterenol, and Galeterone, the level of enzyme inhibition rarely exceeds 90% leaving behind a significant depot for androgen biosynthesis within the tumor. For DHEA-SO₄ to be utilized there is uptake by organic anion transporter polypeptides, deconjugation catalyzed by steroid sulfatase, and adaptive upregulation of prostate steroidogenic enzymes that will convert DHEA into either testosterone or dihydrotestosterone. The depot of DHEA-SO₄ that remains after P450c17 inhibition and the adaptive responses that occur within the tumor to promote DHEA utilization contribute to mechanisms of drug resistance observed with P450c17 inhibitors. Knowledge of these mechanisms identify new targets for therapeutics that could be used to surmount drug resistance in prostate cancer.
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Overall, the specific role of 3β-HSD activity in adipose tissue steroid homeostasis remains to be formally established. Steroid sulfatase (STS) converts DHEA-sulfate (DHEA-S) and estrone-sulfate (E1-S) into their free forms, DHEA and E1 [212]. While it is uncertain that adipose tissue generates de novo DHEA and E1 directly from cholesterol, the sulfated forms of these steroids are highly abundant in the circulation and may represent a significant source for these steroids in adipose tissue, provided that STS is indeed, active [213].
Over the past decade, adipose tissues have been increasingly known for their endocrine properties, that is, their ability to secrete a number of adipocytokines that may exert local and/or systemic effects. In addition, adipose tissues have long been recognized as significant sites for steroid hormone transformation and action. We hereby provide an updated survey of the many steroid-converting enzymes that may be detected in human adipose tissues, their activities and potential roles. In addition to the now well-established role of aromatase and 11β-hydroxysteroid dehydrogenase (HSD) type 1, many enzymes have been reported in adipocyte cell lines, isolated mature cells and/or preadipocytes. These include 11β-HSD type 2, 17β-HSDs, 3β-HSD, 5α-reductases, sulfatases and glucuronosyltransferases. Some of these enzymes are postulated to bear relevance for adipose tissue physiology and perhaps for the pathophysiology of obesity. This elaborate set of steroid-converting enzymes in the cell types of adipose tissue deserves further scientific attention. Our work on 20α-HSD (AKR1C1), 3α-HSD type 3 (AKR1C2) and 17β-HSD type 5 (AKR1C3) allowed us to clarify the relevance of these enzymes for some aspects of adipose tissue function. For example, down-regulation of AKR1C2 expression in preadipocytes seems to potentiate the inhibitory action of dihydrotestosterone on adipogenesis in this model. Many additional studies are warranted to assess the impact of intra-adipose steroid hormone conversions on adipose tissue functions and chronic conditions such as obesity, diabetes and cancer.
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Citation Excerpt :
Since STS possesses broad substrate specificity, it is able to convert CholS, DHEAS and E1S into cholesterol, DHEA and E1 respectively. The use of inhibitors of STS to lower the concentration of active oestrogen in oestrogen-sensitive diseases should take into account the possible side effects due to the inhibition of DHT biosynthesis from ADTS and DHEAS and the accumulation of CholS that causes ichthyosis similar to that found in patients having X-linked ichthyosis due to STS deficiency [62,63]. Cancer cells are cells that have their regulatory machinery altered making them grow and divide in an uncontrolled manner.
There is some confusion in the literature about steroidogenesis in endocrine glands and steroidogenesis in peripheral intracrine tissues. The objective of the present review is to bring some clarifications and better understanding about steroidogenesis in these two types of tissues. Concerns about substrate specificity, kinetic constants and place of enzymes in the pathway have been discussed. The role of 17α-hydroxylase/17–20 lyase (CYP17A1) in the production of dehydroepiandrosterone and back-door pathways of dihydrotestosterone biosynthesis is also analyzed.
This article is part of a Special Issue entitled “Synthesis and biological testing of steroid derivatives as inhibitors”.

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Cell

ArticleCloning and expression of steroid sulfatase cDNA and the frequent occurrence of deletions in STS deficiency: Implications for X-Y interchange

Abstract

Anal. Biochem.

Cell

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Meth. Enzymol.

J. Biol. Chem.

Biochim. Biophys. Acta

Cell

Lancet

J. Mol. Biol.

Cell

Biochim. Biophys. Acta

Variable transfer of Y-specific sequences in XX males

Nucl. Acids Res.

Chromosome Y-specific DNA is transferred to the short arm of X chromosome in human XX males

Science

Screening λgt recombinant clones by hybridization to single plaques in situ

Science

Increased cholesterol sulfate in plasma and red blood cell membranes of steroid sulfatase deficient patients

J. Clin. Endocrinol. Metab.

Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination

Nucleotide sequence of 3-hydroxy-3-methylglutaryl CoA reductase, a glycoprotein of the endoplasmic reticulum

Nature

Inherited chondrodysplasia punctata due to a deletion of the terminal short arm of an X chromosome

N. Eng. J. Med.

Small deletions of the short arm of the Y chromosome in 46, XY females

Recessive X-linked ichthyosis: lack of immunologically detectable steroid sulfatase enzyme protein

Hum. Genet.

Fine mapping of the distal short arm of the human X chromosome using XY translocations

Am. J. Hum. Genet.

A pseudoautosomal gene in man

Science

High frequency of unequal recombination in pseudoautosomal region shown by proviral insertion in transgenic mouse

Nature

X-linkage of steroid sulfatase in the mouse is evidence for a functional Y-linked allele

Nature

Linkage of the murine steroid sulfatase locus and the sex reversed mutation

Am. J. Hum. Genet.

Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumour virus chimaeric plasmids

Nature

Lipids in the pathogenesis of ichthyosis: topical cholesterol sulfate-induced scaling in hairless mice

J. Invest. Dermatol.

Article
Cloning and expression of steroid sulfatase cDNA and the frequent occurrence of deletions in STS deficiency: Implications for X-Y interchange

Buffer gradient gels and ³⁵S label as an aid to rapid DNA sequence determination

Fine mapping of the distal short arm of the human X chromosome using $X Y$ translocations