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Screening of MAMLD1 mutations in 70 children with 46,XY DSD: identification and functional analysis of two new mutations.

Kalfa N, Fukami M, Philibert P, Audran F, Pienkowski C, Weill J, Pinto G, Manouvrier S, Polak M, Ogata T, Sultan C - PLoS ONE (2012)

Bottom Line: The in vitro functional study confirmed no residual transactivating function of the p.S143X mutant and a significantly reduced transactivation function of the p.P384L protein (p = 0.0032).The p.P359S, p.N662S and p.H347Q variants are also reported with particularly high frequency of the p.359T- p.662G haplotype in the DSD patients.Severe undervirilization in XY newborns can reveal mutations of MAMLD1.MAMLD1 should be routinely sequenced in these patients with otherwise normal AR, SRD5A2 and NR5A1genes.

View Article: PubMed Central - PubMed

Affiliation: Service d'Hormonologie, Hôpital Lapeyronie, CHU de Montpellier et UM1, Montpellier, France.

ABSTRACT
More than 50% of children with severe 46,XY disorders of sex development (DSD) do not have a definitive etiological diagnosis. Besides gonadal dysgenesis, defects in androgen biosynthesis, and abnormalities in androgen sensitivity, the Mastermind-like domain containing 1 (MAMLD1) gene, which was identified as critical for the development of male genitalia, may be implicated. The present study investigated whether MAMLD1 is implicated in cases of severe 46,XY DSD and whether routine sequencing of MAMLD1 should be performed in these patients.Seventy children with severe non-syndromic 46,XY DSD of unknown etiology were studied. One hundred and fifty healthy individuals were included as controls. Direct sequencing of the MAMLD1, AR, SRD5A2 and NR5A1 genes was performed. The transactivation function of the variant MAMLD1 proteins was quantified by the luciferase method.TWO NEW MUTATIONS WERE IDENTIFIED: p.S143X (c.428C>A) in a patient with scrotal hypospadias with microphallus and p.P384L (c.1151C>T) in a patient with penile hypospadias with microphallus. The in vitro functional study confirmed no residual transactivating function of the p.S143X mutant and a significantly reduced transactivation function of the p.P384L protein (p = 0.0032). The p.P359S, p.N662S and p.H347Q variants are also reported with particularly high frequency of the p.359T- p.662G haplotype in the DSD patients.Severe undervirilization in XY newborns can reveal mutations of MAMLD1. MAMLD1 should be routinely sequenced in these patients with otherwise normal AR, SRD5A2 and NR5A1genes.

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Related in: MedlinePlus

Tertiary structure prediction of the wildtype protein (left column) and with the mutants.3D structure was predicted at Protein Homology/analogY Recognition Engine (PhyreEngine) from the Structural Bioinformatics Group, Imperial College, London, at http:www.sbg.bio.ic.ac.uk/phyre~/. The plain arrows show the changes in the shape of the protein between the wildtype and p.P384L.
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pone-0032505-g002: Tertiary structure prediction of the wildtype protein (left column) and with the mutants.3D structure was predicted at Protein Homology/analogY Recognition Engine (PhyreEngine) from the Structural Bioinformatics Group, Imperial College, London, at http:www.sbg.bio.ic.ac.uk/phyre~/. The plain arrows show the changes in the shape of the protein between the wildtype and p.P384L.

Mentions: a- The p.S143X mutation was predicted to cause a short and truncated protein. The in silico prediction showed profoundly modified amino acid accessibility and 3D structure. Relative surface accessibility and absolute surface accessibility of the last amino acid changed from 0.248 to 0.834 and from 29.124 to 97.721, respectively. PhyreEngine predicted the loss of any functional site without a residual consensus sequence (no homologous sequence over 5% through whole genome) (Fig. 2). The in vitro functional study confirmed no residual transactivating function of the mutant (Fig. 3). Interestingly, a maternal uncle and a maternal cousin of the index case both exhibited severe hypospadias (not available for genetic testing). The mother was indeed heterozygous for the mutation (Fig. 1).


Screening of MAMLD1 mutations in 70 children with 46,XY DSD: identification and functional analysis of two new mutations.

Kalfa N, Fukami M, Philibert P, Audran F, Pienkowski C, Weill J, Pinto G, Manouvrier S, Polak M, Ogata T, Sultan C - PLoS ONE (2012)

Tertiary structure prediction of the wildtype protein (left column) and with the mutants.3D structure was predicted at Protein Homology/analogY Recognition Engine (PhyreEngine) from the Structural Bioinformatics Group, Imperial College, London, at http:www.sbg.bio.ic.ac.uk/phyre~/. The plain arrows show the changes in the shape of the protein between the wildtype and p.P384L.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316539&req=5

pone-0032505-g002: Tertiary structure prediction of the wildtype protein (left column) and with the mutants.3D structure was predicted at Protein Homology/analogY Recognition Engine (PhyreEngine) from the Structural Bioinformatics Group, Imperial College, London, at http:www.sbg.bio.ic.ac.uk/phyre~/. The plain arrows show the changes in the shape of the protein between the wildtype and p.P384L.
Mentions: a- The p.S143X mutation was predicted to cause a short and truncated protein. The in silico prediction showed profoundly modified amino acid accessibility and 3D structure. Relative surface accessibility and absolute surface accessibility of the last amino acid changed from 0.248 to 0.834 and from 29.124 to 97.721, respectively. PhyreEngine predicted the loss of any functional site without a residual consensus sequence (no homologous sequence over 5% through whole genome) (Fig. 2). The in vitro functional study confirmed no residual transactivating function of the mutant (Fig. 3). Interestingly, a maternal uncle and a maternal cousin of the index case both exhibited severe hypospadias (not available for genetic testing). The mother was indeed heterozygous for the mutation (Fig. 1).

Bottom Line: The in vitro functional study confirmed no residual transactivating function of the p.S143X mutant and a significantly reduced transactivation function of the p.P384L protein (p = 0.0032).The p.P359S, p.N662S and p.H347Q variants are also reported with particularly high frequency of the p.359T- p.662G haplotype in the DSD patients.Severe undervirilization in XY newborns can reveal mutations of MAMLD1.MAMLD1 should be routinely sequenced in these patients with otherwise normal AR, SRD5A2 and NR5A1genes.

View Article: PubMed Central - PubMed

Affiliation: Service d'Hormonologie, Hôpital Lapeyronie, CHU de Montpellier et UM1, Montpellier, France.

ABSTRACT
More than 50% of children with severe 46,XY disorders of sex development (DSD) do not have a definitive etiological diagnosis. Besides gonadal dysgenesis, defects in androgen biosynthesis, and abnormalities in androgen sensitivity, the Mastermind-like domain containing 1 (MAMLD1) gene, which was identified as critical for the development of male genitalia, may be implicated. The present study investigated whether MAMLD1 is implicated in cases of severe 46,XY DSD and whether routine sequencing of MAMLD1 should be performed in these patients.Seventy children with severe non-syndromic 46,XY DSD of unknown etiology were studied. One hundred and fifty healthy individuals were included as controls. Direct sequencing of the MAMLD1, AR, SRD5A2 and NR5A1 genes was performed. The transactivation function of the variant MAMLD1 proteins was quantified by the luciferase method.TWO NEW MUTATIONS WERE IDENTIFIED: p.S143X (c.428C>A) in a patient with scrotal hypospadias with microphallus and p.P384L (c.1151C>T) in a patient with penile hypospadias with microphallus. The in vitro functional study confirmed no residual transactivating function of the p.S143X mutant and a significantly reduced transactivation function of the p.P384L protein (p = 0.0032). The p.P359S, p.N662S and p.H347Q variants are also reported with particularly high frequency of the p.359T- p.662G haplotype in the DSD patients.Severe undervirilization in XY newborns can reveal mutations of MAMLD1. MAMLD1 should be routinely sequenced in these patients with otherwise normal AR, SRD5A2 and NR5A1genes.

Show MeSH
Related in: MedlinePlus