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A Novel Mutation of DAX-1 Associated with Secretory Azoospermia.

Mou L, Xie N, Yang L, Liu Y, Diao R, Cai Z, Li H, Gui Y - PLoS ONE (2015)

Bottom Line: A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified.Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1.This novel mutation (p.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Genitourinary Tumor, Shenzhen Domesticated Organ Medical Engineering Research and Development Center, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China; Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Biomedical Research Institute, Shenzhen PKU-HKUST Medical Center, Shenzhen, 518036, China.

ABSTRACT
Secretory azoospermia is a severe form of male infertility caused by unknown factors. DAX-1 is predominantly expressed in mammalian reproductive tissues and plays an important role in spermatogenesis because Dax-1 knockout male mice show spermatogenesis defects. To examine whether DAX-1 is involved in the pathogenesis of secretory azoospermia in humans, we sequenced all of the exons of DAX-1 in 776 patients diagnosed with secretory azoospermia and 709 proven fertile men. A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified. Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1. This novel mutation (p. V385L) of DAX-1 is the first to be identified in association with secretory azoospermia, thereby highlighting the important role of DAX-1 in spermatogenesis.

No MeSH data available.


Related in: MedlinePlus

Six missense mutations in DAX-1 identified in patients with secretory azoospermia.(A) Chromatogram traces from Sanger sequencing, showing the validated missense mutations. (B) Evolutionary conservation of amino acids affected by the missense mutations. Multiple protein alignments were performed with MegAlign (Demonstration System DNASTAR, Inc.). The identification numbers of the DAX-1 protein were as follows: human (NP_000466.2), chimpanzee (XP_520991.2), rhesus (XP_002806222.1), pig (NP_999552.1), rat (NP_445769.1), mouse (NP_031456.1), chicken (NP_989924.1), and Medaka fish (NP_001104259.1). The mutant alleles are boxed, and the star (*) indicates the conserved residue.
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pone.0133997.g001: Six missense mutations in DAX-1 identified in patients with secretory azoospermia.(A) Chromatogram traces from Sanger sequencing, showing the validated missense mutations. (B) Evolutionary conservation of amino acids affected by the missense mutations. Multiple protein alignments were performed with MegAlign (Demonstration System DNASTAR, Inc.). The identification numbers of the DAX-1 protein were as follows: human (NP_000466.2), chimpanzee (XP_520991.2), rhesus (XP_002806222.1), pig (NP_999552.1), rat (NP_445769.1), mouse (NP_031456.1), chicken (NP_989924.1), and Medaka fish (NP_001104259.1). The mutant alleles are boxed, and the star (*) indicates the conserved residue.

Mentions: To examine whether DAX-1 genetic defects are associated with secretory azoospermia, we screened for DAX-1 exonic mutations in 776 secretory azoospermia patients and 709 men with proven fertility using massively parallel sequencing technology. As shown in Table 1, six missense mutations and six synonymous mutations were detected in DAX-1. All of the mutations except for c.498 G>A, c.376 G>A and c.114 C>T were not present in either dbSNP135 database or 1000 Genome Project dataset and were not identified in the 709 normal controls. The six novel missense mutations (c.152 G>A, c.321 C>G, c.725 C>T, c.766 G>C, c.1153 G>T and c.1279 A>G) were further confirmed by Sanger sequencing (Fig 1A). Alignment of the amino acid sequence of DAX-1 to its orthologs in different species showed that the V385L mutation affected a highly conserved amino acid (Fig 1B). Bioinformatic assessment of the variants indicated that the mutations C104W, E256Q and V385L were possibly damaging to the protein predicted by both Polyphen 2.0[24] and MutationTaster[25] (Table 2).


A Novel Mutation of DAX-1 Associated with Secretory Azoospermia.

Mou L, Xie N, Yang L, Liu Y, Diao R, Cai Z, Li H, Gui Y - PLoS ONE (2015)

Six missense mutations in DAX-1 identified in patients with secretory azoospermia.(A) Chromatogram traces from Sanger sequencing, showing the validated missense mutations. (B) Evolutionary conservation of amino acids affected by the missense mutations. Multiple protein alignments were performed with MegAlign (Demonstration System DNASTAR, Inc.). The identification numbers of the DAX-1 protein were as follows: human (NP_000466.2), chimpanzee (XP_520991.2), rhesus (XP_002806222.1), pig (NP_999552.1), rat (NP_445769.1), mouse (NP_031456.1), chicken (NP_989924.1), and Medaka fish (NP_001104259.1). The mutant alleles are boxed, and the star (*) indicates the conserved residue.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4514677&req=5

pone.0133997.g001: Six missense mutations in DAX-1 identified in patients with secretory azoospermia.(A) Chromatogram traces from Sanger sequencing, showing the validated missense mutations. (B) Evolutionary conservation of amino acids affected by the missense mutations. Multiple protein alignments were performed with MegAlign (Demonstration System DNASTAR, Inc.). The identification numbers of the DAX-1 protein were as follows: human (NP_000466.2), chimpanzee (XP_520991.2), rhesus (XP_002806222.1), pig (NP_999552.1), rat (NP_445769.1), mouse (NP_031456.1), chicken (NP_989924.1), and Medaka fish (NP_001104259.1). The mutant alleles are boxed, and the star (*) indicates the conserved residue.
Mentions: To examine whether DAX-1 genetic defects are associated with secretory azoospermia, we screened for DAX-1 exonic mutations in 776 secretory azoospermia patients and 709 men with proven fertility using massively parallel sequencing technology. As shown in Table 1, six missense mutations and six synonymous mutations were detected in DAX-1. All of the mutations except for c.498 G>A, c.376 G>A and c.114 C>T were not present in either dbSNP135 database or 1000 Genome Project dataset and were not identified in the 709 normal controls. The six novel missense mutations (c.152 G>A, c.321 C>G, c.725 C>T, c.766 G>C, c.1153 G>T and c.1279 A>G) were further confirmed by Sanger sequencing (Fig 1A). Alignment of the amino acid sequence of DAX-1 to its orthologs in different species showed that the V385L mutation affected a highly conserved amino acid (Fig 1B). Bioinformatic assessment of the variants indicated that the mutations C104W, E256Q and V385L were possibly damaging to the protein predicted by both Polyphen 2.0[24] and MutationTaster[25] (Table 2).

Bottom Line: A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified.Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1.This novel mutation (p.

View Article: PubMed Central - PubMed

Affiliation: Shenzhen Key Laboratory of Genitourinary Tumor, Shenzhen Domesticated Organ Medical Engineering Research and Development Center, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, 518035, China; Guangdong and Shenzhen Key Laboratory of Male Reproductive Medicine and Genetics, Institute of Urology, Peking University Shenzhen Hospital, Biomedical Research Institute, Shenzhen PKU-HKUST Medical Center, Shenzhen, 518036, China.

ABSTRACT
Secretory azoospermia is a severe form of male infertility caused by unknown factors. DAX-1 is predominantly expressed in mammalian reproductive tissues and plays an important role in spermatogenesis because Dax-1 knockout male mice show spermatogenesis defects. To examine whether DAX-1 is involved in the pathogenesis of secretory azoospermia in humans, we sequenced all of the exons of DAX-1 in 776 patients diagnosed with secretory azoospermia and 709 proven fertile men. A number of coding mutations unique to the patient group, including two synonymous mutations and six missense mutations, were identified. Of the missense mutations, our functional assay demonstrated that the V385L mutation caused the reduced functioning of DAX-1. This novel mutation (p. V385L) of DAX-1 is the first to be identified in association with secretory azoospermia, thereby highlighting the important role of DAX-1 in spermatogenesis.

No MeSH data available.


Related in: MedlinePlus