title: Identification of a new DPY19L2 mutation and a better definition of DPY19L2 deletion breakpoints leading to globozoospermia.
abstract: OBJECTIVE
  The purpose of this study was to analyze DPY19L2 sequence variants to investigate the mechanism leading to the entire DPY19L2 deletion in a large cohort of infertile globozoospermic patients.
  
  
  RESULTS
  An improved analysis of the DPY19L2 deletion breakpoints (BPs) allowed us to identify two BPs located in a small 1 kb region and to more precisely localize the BPs reported previously.
  
  
  BACKGROUND
  Three genes [spermatogenesis associated 16 (SPATA16), protein interacting with PRKCA (PICK1) and DPY19L2] were previously correlated with globozoospermia, but a homozygous deletion of the entire DPY19L2 was identified as the most frequent alteration causing this phenotype. In addition, several point mutations in this gene were reported. In previous work, we have identified nine BPs for the DPY19L2 deletion clustered in two hotspot regions, while others reported a total of five BPs.
  
  
  METHODS
  We screened for the DPY19L2 deletion and for mutations in the DPY19L2, SPATA16 and PICK1 genes in a cohort of 21 Tunisian globozoospermic patients. In order to characterize the DPY19L2 deletion BPs, we sequenced a 2 kb fragment on low copy repeat (LCR) 1 and LCR2 in Tunisian fertile controls to distinguish between single-nucleotide polymorphisms (SNPs) and LCR-specific markers.
  
  
  RESULTS
  Molecular analyses performed on 18 genetically independent individuals showed that 11 (61.1%) were homozygous for the DPY19L2 deletion, 2 (11.1%) were homozygous for the non-synonymous mutation (p.R298C) in exon 8, 1 patient (5.6%) was homozygous for a new splice-site mutation at the junction exon-intron 16 [c.1579_1580+4delAGGTAAinsTCAT] and no DPY19L2, SPATA16 or PICK1 mutations were identified for 4 patients (22.2%). By defining 15 specific LCR markers, we characterized 2 BPs for the DPY19L2 deletion in 11 patients showing the homozygous deletion. Using 20 non-LCR-specific SNPs, we identified 8 distinct haplotypes.
  
  
  CONCLUSIONS
  A limitation of this study is the small number of patients owing to the rarity of this form of male infertility.
  
  
  CONCLUSIONS
  Our data showed that some nucleotides, described by others as LCR-specific markers and used to limit their BPs, were in fact SNPs demonstrating the difficulty in precisely determining the localization of BPs.
  
  
  METHODS
  Not applicable.
  
  
  BACKGROUND
  This work was supported by the French Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM), the Ministère de l'Education Nationale et de l'Enseignement Supérieur et de la Recherche, the University of Strasbourg, the University Hospital of Strasbourg, the Agence Nationale pour la Recherche, the Agence de la BioMédecine and l'Agence Universitaire de la Francophonie (AUF). There are no conflicts of interest to declare.
authors.affiliation: Laboratoire de Cytogénétique, Génétique Moléculaire et Biologie de la Reproduction Humaines, CHU Farhat Hached, 4000 Sousse, Tunisia.
authors.forename: Houda
authors.initials: H
authors.lastname: Ghédir
authors.affiliation: Laboratoire de Cytogénétique, Génétique Moléculaire et Biologie de la Reproduction Humaines, CHU Farhat Hached, 4000 Sousse, Tunisia.
authors.forename: Samira
authors.initials: S
authors.lastname: Ibala-Romdhane
authors.affiliation: Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC), Institut National de Santé et de Recherche Médicale INSERM U964, Centre National de Recherche scientifique CNRS UMR 1704, Université de Strasbourg, Illkirch 67404, France Centre Hospitalier Universitaire, Strasbourg F-67000, France.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Okutman
authors.affiliation: Unité de Génétique Médicale, Maternité Port-Royal, Hôpital Cochin, 75679 Paris 14, France.
authors.forename: Géraldine
authors.initials: G
authors.lastname: Viot
authors.affiliation: Laboratoire de Cytogénétique, Génétique Moléculaire et Biologie de la Reproduction Humaines, CHU Farhat Hached, 4000 Sousse, Tunisia.
authors.forename: Ali
authors.initials: A
authors.lastname: Saad
authors.affiliation: Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC), Institut National de Santé et de Recherche Médicale INSERM U964, Centre National de Recherche scientifique CNRS UMR 1704, Université de Strasbourg, Illkirch 67404, France Centre Hospitalier Universitaire, Strasbourg F-67000, France viville@igbmc.fr.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Viville
mesh_terms: D000177:Acrosome / Q000648:ultrastructure; D000483:Alleles; D002352:Carrier Proteins / Q000235:genetics; D056905:Chromosome Breakpoints; D003241:Consanguinity; D005091:Exons / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage; D006239:Haplotypes / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D007248:Infertility, Male / Q000235:genetics*; D008297:Male; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics* / Q000502:physiology; D009687:Nuclear Proteins / Q000235:genetics; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D022821:RNA Splice Sites / Q000235:genetics; D016415:Sequence Alignment; D013094:Spermatozoa / Q000002:abnormalities / Q000648:ultrastructure; D014416:Tunisia / Q000453:epidemiology; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; C511317:DPY19L2 protein, human; D018398:Homeodomain Proteins; D008565:Membrane Proteins; D009687:Nuclear Proteins; C498297:PICk1 protein, human; D022821:RNA Splice Sites; C474264:SPATA16 protein, human; D033921:Vesicular Transport Proteins
keywords: DPY19L2; breakpoints; globozoospermia; male infertility; non-allelic homologous recombination; round-headed