Functional study of a novel c.630delG (p.Y211Tfs*85) mutation in NR5A1 gene in a Chinese boy with 46,XY disorders of sex development

  • Sinian PanEmail author
  • Shili Guo
  • Liting Liu
  • Xiaoyuan Yang
  • Hanmei Liang



This study aimed to present the clinical features and gene mutation characteristics of a child with 46,XY disorders of sex development (DSD) caused by a novel heterozygous mutation in the NR5A1 gene to determine the potential association between this heterozygous mutation and the pathogenesis of 46,XY DSD.


We present the case of a Chinese child with ambiguous genitalia at birth but a normal adrenal gland. Targeted next-generation sequencing, comprising 163 candidate genes involved in sexual differentiation and development, was performed, followed by the functional evaluation of the novel NR5A1 mutation.


The patient had a novel heterozygous mutation in the NR5A1 gene, c.630delG (p.Y211Tfs*85). Results revealed that overexpression of p.Y211Tfs*85 impaired steroidogenic factor-1 (SF-1) protein synthesis. Immunofluorescence analysis revealed that both SF-1 wild-type and p.Y211Tfs*85 mutation proteins were localized in the cell nucleus. Furthermore, dual-luciferase reporter assay results revealed that the p.Y211Tfs*85 mutation could effectively downregulate the transcriptional activation of anti-Müllerian hormone and steroidogenic acute regulatory protein genes (P < 0.01). Additionally, the p.Y211Tfs*85 mutation changed three-dimensional conformation of SF-1, and three conformations could be constructed with the mutated amino acid sequences. Therefore, the novel frameshift mutation could result in decreased protein expression of SF-1.


We described a novel mutation in NR5A1 and showed that it might affect protein structure, thereby seriously compromising the role of SF-1 in regulating gonadal development. The novel p.Y211Tfs*85 mutation in the NR5A1 gene enriches the boy of information available regarding the mutation spectrum of this gene in the Chinese population.


NR5A1 SF-1 AMH STAR Disorders of sex development 


Authors’ contributions

• Sinian Pan and Shili Guo: Formal analysis, funding acquisition, investigation,; methodology, writing original draft

• Liting Liu: Supervision, validation

• Xiaoyuan Yang and Hanmei Liang: Review and editing

Funding information

This work was supported by the Guangdong Science and Technology project (grant number 2012B031800077).

Compliance with ethical standards

Ethics statement

The Research Ethics Committee of the Third Affiliated Hospital of Sun Yat-sen University approved the collection of tissue samples for research.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Pasterski V, Prentice P, Hughes IA. Impact of the consensus statement and the new DSD classification system. Best Pract Res Clin Endocrinol Metab. 2010;24(2):187–95. Scholar
  2. 2.
    Nagy O, Karteszi J, Hartwig M, Bertalan R, Javorszky E, Erhardt E, et al. The importance of the multiplex ligation-dependent probe amplification in the identification of a novel two-exon deletion of the NR5A1 gene in a patient with 46,XY differences of sex development. Mol Biol Rep. 2019;46:5595–601. Scholar
  3. 3.
    Werner R, Monig I, Lunstedt R, Wunsch L, Thorns C, Reiz B, et al. New NR5A1 mutations and phenotypic variations of gonadal dysgenesis. PLoS One. 2017;12(5):e0176720. Scholar
  4. 4.
    Anamthathmakula P, Miryala CSJ, Moreci RS, Kyathanahalli C, Hassan SS, Condon JC, et al. Steroidogenic factor 1 (Nr5a1) is required for Sertoli cell survival post sex determination. Sci Rep. 2019;9(1):4452. Scholar
  5. 5.
    Lin L, Achermann JC. Steroidogenic factor-1 (SF-1, Ad4BP, NR5A1) and disorders of testis development. Sexual Development : genetics, molecular biology, evolution, endocrinology, embryology, and pathology of sex determination and differentiation. 2008;2(4–5):200–9. Scholar
  6. 6.
    Suntharalingham JP, Buonocore F, Duncan AJ, Achermann JC. DAX-1 (NR0B1) and steroidogenic factor-1 (SF-1, NR5A1) in human disease. Best Pract Res Clin Endocrinol Metab. 2015;29(4):607–19. Scholar
  7. 7.
    Sekido R, Lovell-Badge R. Sex determination involves synergistic action of SRY and SF1 on a specific Sox9 enhancer. Nature. 2008;453(7197):930–4. Scholar
  8. 8.
    Bashamboo A, McElreavey K. Human sex-determination and disorders of sex-development (DSD). Semin Cell Dev Biol. 2015;45:77–83. Scholar
  9. 9.
    Fabbri HC, Soardi FC, Calais FLD, Petroli RJ, Maciel-Guerra AT, Guerra-Júnior G et al. The novel p.Cys65Tyr mutation in NR5A1 gene in three 46,XY siblings with normal testosterone levels and their mother with primary ovarian insufficiency. BMC Med Genet,15,1(2014-01-10). 2014;15(1):7.Google Scholar
  10. 10.
    Swartz JM, Ciarlo R, Guo MH, Abrha A, Weaver B, Diamond DA, et al. A 46,XX ovotesticular disorder of sex development likely caused by a steroidogenic factor-1 (NR5A1) variant. Horm Res Paediatr. 2017;87(3):191–5. Scholar
  11. 11.
    Granados A, Alaniz VI, Mohnach L, Barseghyan H, Vilain E, Ostrer H, et al. MAP3K1-related gonadal dysgenesis: six new cases and review of the literature. Am J Med Genet C: Semin Med Genet. 2017;175(2):253–9. Scholar
  12. 12.
    Bashamboo A, Donohoue PA, Vilain E, Rojo S, Calvel P, Seneviratne SN, et al. A recurrent p.Arg92Trp variant in steroidogenic factor-1 (NR5A1) can act as a molecular switch in human sex development. Hum Mol Genet. 2016;25(23):5286. Scholar
  13. 13.
    Kohler B, Lin L, Ferraz-de-Souza B, Wieacker P, Heidemann P, Schroder V, et al. Five novel mutations in steroidogenic factor 1 (SF1, NR5A1) in 46,XY patients with severe underandrogenization but without adrenal insufficiency. Hum Mutat. 2008;29(1):59–64. Scholar
  14. 14.
    Baetens D, Stoop H, Peelman F, Todeschini AL, Rosseel T, Coppieters F, et al. NR5A1 is a novel disease gene for 46,XX testicular and ovotesticular disorders of sex development. Genet Med. 2017;19(4):367–76. Scholar
  15. 15.
    Robevska G, van den Bergen JA, Ohnesorg T, Eggers S, Hanna C, Hersmus R, et al. Functional characterization of novel NR5A1 variants reveals multiple complex roles in disorders of sex development. Hum Mutat. 2018;39(1):124–39. Scholar
  16. 16.
    Fabbri-Scallet H, de Mello MP, Guerra-Junior G, Maciel-Guerra AT, de Andrade JGR, de Queiroz CMC, et al. Functional characterization of five NR5A1 gene mutations found in patients with 46,XY disorders of sex development. Hum Mutat. 2018;39(1):114–23. Scholar
  17. 17.
    Arnold K, Bordoli L, Kopp J, Schwede T. The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics (Oxford, England). 2006;22(2):195–201. Scholar
  18. 18.
    Achermann JC, Ito M, Ito M, Hindmarsh PC, Jameson JL. A mutation in the gene encoding steroidogenic factor-1 causes XY sex reversal and adrenal failure in humans. Nat Genet. 1999;22(2):125–6. Scholar
  19. 19.
    Lin L, Philibert P, Ferraz-de-Souza B, Kelberman D, Homfray T, Albanese A, et al. Heterozygous missense mutations in steroidogenic factor 1 (SF1/Ad4BP, NR5A1) are associated with 46,XY disorders of sex development with normal adrenal function. J Clin Endocrinol Metab. 2007;92(3):991–9. Scholar
  20. 20.
    Kohler B, Lin L, Mazen I, Cetindag C, Biebermann H, Akkurt I, et al. The spectrum of phenotypes associated with mutations in steroidogenic factor 1 (SF-1, NR5A1, Ad4BP) includes severe penoscrotal hypospadias in 46,XY males without adrenal insufficiency. Eur J Endocrinol. 2009;161(2):237–42. Scholar
  21. 21.
    Ferraz-de-Souza B, Lin L, Achermann JC. Steroidogenic factor-1 (SF-1, NR5A1) and human disease. Mol Cell Endocrinol. 2011;336(1–2):198–205. Scholar
  22. 22.
    Luo X, Ikeda Y, Parker KL. A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differentiation. Cell. 1994;77(4):481–90. Scholar
  23. 23.
    Tremblay JJ, Viger RS. A mutated form of steroidogenic factor 1 (SF-1 G35E) that causes sex reversal in humans fails to synergize with transcription factor GATA-4. J Biol Chem. 2003;278(43):42637–42. Scholar
  24. 24.
    Harrison SM, Campbell IM, Keays M, Granberg CF, Villanueva C, Tannin G, et al. Screening and familial characterization of copy-number variations in NR5A1 in 46,XY disorders of sex development and premature ovarian failure. Am J Med Genet A. 2013;161a(10):2487–94. Scholar
  25. 25.
    Fabbri HC, de Andrade JG, Soardi FC, de Calais FL, Petroli RJ, Maciel-Guerra AT, et al. The novel p.Cys65Tyr mutation in NR5A1 gene in three 46,XY siblings with normal testosterone levels and their mother with primary ovarian insufficiency. BMC Med Genet. 2014;15:7. Scholar
  26. 26.
    Zangen D, Kaufman Y, Zeligson S, Perlberg S, Fridman H, Kanaan M, et al. XX ovarian dysgenesis is caused by a PSMC3IP/HOP2 mutation that abolishes coactivation of estrogen-driven transcription. Am J Hum Genet. 2011;89(4):572–9. Scholar
  27. 27.
    Kalfa N, Fukami M, Philibert P, Audran F, Pienkowski C, Weill J, et al. Screening of MAMLD1 mutations in 70 children with 46,XY DSD: identification and functional analysis of two new mutations. PLoS One. 2012;7(3):e32505. Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.The Department of PediatricsThe Third Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina

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