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Updated: Sep 25, 2025
A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
Published on: September 15, 2021
Identification of a new splice-acceptor mutation in HFM1 and functional analysis through molecular docking in
Neda Saebnia1, Reza Ebrahimzadeh-Vesal2, Aliakbar Haddad-Mashhadrizeh3
1Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Purpose:
To investigate the genetic cause of nonobstructive azoospermia (NOA).
Methods:
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.
Results:
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.
Conclusion:
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.
