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Identification of a new splice-acceptor mutation in HFM1 and functional analysis through molecular docking in nonobstructive azoospermia



To investigate the genetic cause of nonobstructive azoospermia (NOA).


We performed whole exome sequencing (WES) on the proband who had three relatives suffering from NOA. We used a list of candidate genes which have high expression level in testis and their mutations have been reported in NOA. Sanger sequencing verified the identified variant and its structural and functional consequence was evaluated by protein three-dimensional (3D) structure prediction and protein-ligand docking.


WES revealed a novel splice-acceptor mutation (c.1832-2A>T) in helicase for meiosis 1 (HFM1) gene, which co-segregated with the NOA in this family. 3D structural models were generated and verified. Molecular docking indicated that the c.1832-2A>T mutation affects not only the ADP binding residues but also the hydrogen bond interactions. The ADP binding site will be lost in the mutant protein, potentially causing defective crossover and synapsis.


We report that the c.1832-2A>T mutation is the likely cause of NOA in the family studied. Regarding that many reported NOA genes are involved in the formation of crossovers and synapsis and have critical roles in the production of germ cells, we suggest that such genes should be considered for screening of infertility among large cohorts of infertile individuals.

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The authors thank the patients and their family members who participated in this study. We appreciate the contribution of Ferdowsi University of Mashhad and Neyshabur University of Medical Sciences to provide facilities to accomplish this study. We also appreciate Novin Fertility Center for introducing patients to our study. We gratefully acknowledge Professor Pierre F. Ray and Dr. Reza Maroofian for the financial support, Nicolas Thierry-Mieg for his support in bioinformatics concerns, and Abolfazl Rad for his help in extraction and sending DNA samples for WES. The authors thank the management of advanced computational center, Khayyam Innovation Ecosystem, Mashhad, Iran for providing the facilities and encouragement to carry out this research work.

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Correspondence to Zeinab Neshati or Mohsen Azimi-Nezhad.

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Saebnia, N., Ebrahimzadeh-Vesal, R., Haddad-Mashhadrizeh, A. et al. Identification of a new splice-acceptor mutation in HFM1 and functional analysis through molecular docking in nonobstructive azoospermia. J Assist Reprod Genet 39, 1195–1203 (2022).

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  • Male infertility
  • Nonobstructive azoospermia
  • Whole exome sequencing
  • Protein modeling
  • Molecular docking
  • HFM1