title: SUMO1 in human sperm: new targets, role in motility and morphology and relationship with DNA damage.
abstract: In studies carried out previously, we demonstrated that small ubiquitin-like modifier 1 (SUMO1) is associated with poor sperm motility when evaluated with a protocol that reveals mostly SUMO1-ylated live sperm. Recently, with another protocol, it has been demonstrated that SUMO is expressed in most sperm and is related to poor morphology and motility, suggesting that sumoylation may have multiple roles depending on its localisation and targets. We show herein, by confocal microscopy and co-immunoprecipitation, that dynamin-related protein 1 (DRP1), Ran GTPase-activating protein 1 (RanGAP1) and Topoisomerase IIα, SUMO1 targets in somatic and/or germ cells, are SUMO1-ylated in mature human spermatozoa. DRP1 co-localises with SUMO1 in the mid-piece, whereas RanGAP1 and Topoisomerase IIα in the post-acrosomal region of the head. Both SUMO1 expression and co-localisation with the three proteins were significantly higher in morphologically abnormal sperm, suggesting that sumoylation represents a marker of defective sperm. DRP1 sumoylation at the mid-piece level was higher in the sperm of asthenospermic men. As in somatic cells, DRP1 sumoylation is associated with mitochondrial alterations, this protein may represent the link between SUMO and poor motility. As SUMO pathways are involved in responses to DNA damage, another aim of our study was to investigate the relationship between sumoylation and sperm DNA fragmentation (SDF). By flow cytometry, we demonstrated that SUMO1-ylation and SDF are correlated (r=0.4, P<0.02, n=37) and most sumoylated sperm shows DNA damage in co-localisation analysis. When SDF was induced by stressful conditions (freezing and thawing and oxidative stress), SUMO1-ylation increased. Following freezing and thawing, SUMO1-Topoisomerase IIα co-localisation and co-immunoprecipitation increased, suggesting an involvement in the formation/repair of DNA breakage.
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy elisabetta.baldi@unifi.it sara.marchiani@unifi.it.
authors.forename: S
authors.initials: S
authors.lastname: Marchiani
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Tamburrino
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Ricci
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: D
authors.initials: D
authors.lastname: Nosi
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Cambi
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Piomboni
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Belmonte
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Forti
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Muratori
authors.affiliation: Department of BiomedicalExperimental and Clinical Sciences, Center of Excellence DeNotheDepartment of Experimental and Clinical MedicineUniversity of Florence, Viale Pieraccini 6, I-50139 Florence, ItalyDepartments of Molecular and Developmental MedicineMedicalSurgical and Neurological Sciences, University of Siena, 53100 Siena, Italy elisabetta.baldi@unifi.it sara.marchiani@unifi.it.
authors.forename: E
authors.initials: E
authors.lastname: Baldi
mesh_terms: D000951:Antigens, Neoplasm / Q000378:metabolism; D048430:Cell Shape; D003080:Cold Temperature; D015925:Cryopreservation; D004249:DNA Damage; D053938:DNA Fragmentation; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000378:metabolism; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism / Q000473:pathology; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology; D024101:Mitochondrial Proteins / Q000378:metabolism; D018384:Oxidative Stress; D025842:SUMO-1 Protein / Q000378:metabolism*; D015398:Signal Transduction; D013077:Sperm Head / Q000378:metabolism / Q000473:pathology; D013081:Sperm Motility; D013094:Spermatozoa / Q000378:metabolism* / Q000473:pathology; D058207:Sumoylation
chemical_list: D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; D020690:GTPase-Activating Proteins; D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; C092317:RANGAP1 protein, human; D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins; D004250:DNA Topoisomerases, Type II