Identification and characterization of RING-finger ubiquitin ligase UBR7 in mammalian spermatozoa
| Autor: | Miriam Sutovsky, Fred W. van Leeuwen, Gavin C. Conant, Peter Sutovsky, Shawn Zimmerman, Young-Joo Yi |
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| Přispěvatelé: | Psychiatrie & Neuropsychologie, RS: MHeNs - R3 - Neuroscience |
| Rok vydání: | 2014 |
| Předmět: |
Male
endocrine system UBR7 Histology Mouse Swine Spermiogenesis Ubiquitin-Protein Ligases C14ORF130 Blotting Western Fluorescent Antibody Technique Fertilization in Vitro Biology Acrosomal matrix Pathology and Forensic Medicine Mice Ubiquitin Testis medicine Animals Humans RNA Messenger Ligase activity Antibodies Blocking Zona pellucida Acrosome Phylogeny reproductive and urinary physiology Pig Proteasome Spermatid urogenital system Cell Biology Spermatids Spermatozoa Molecular biology Ubiquitin ligase Protein Transport medicine.anatomical_structure Fertilization biology.protein |
| Zdroj: | Cell and Tissue Research, 356(1), 261-278. Springer Verlag |
| ISSN: | 1432-0878 0302-766X |
| DOI: | 10.1007/s00441-014-1808-x |
| Popis: | The ubiquitin-proteasome system (UPS) controls intracellular protein turnover in a substrate-specific manner via E3-type ubiquitin ligases. Mammalian fertilization and particularly sperm penetration through the oocyte vitelline coat, the zona pellucida (ZP), is regulated by UPS. We use an extrinsic substrate of the proteasome-dependent ubiquitin-fusion degradation pathway, the mutant ubiquitin UBB(+1), to provide evidence that an E3-type ligase activity exists in sperm-acrosomal fractions. Protein electrophoresis gels from such de novo ubiquitination experiments contained a unique protein band identified by tandem mass spectrometry as being similar to ubiquitin ligase UBR7 (alternative name: C14ORF130). Corresponding mRNA was amplified from boar testis and several variants of the UBR7 protein were detected in boar, mouse and human sperm extracts by Western blotting. Genomic analysis indicated a high degree of evolutionary conservation, remarkably constant purifying selection and conserved testis expression of the UBR7 gene. By immunofluorescence, UBR7 was localized to the spermatid acrosomal cap and sperm acrosome, in addition to hotspots of proteasomal activity in spermatids, such as the cytoplasmic lobe, caudal manchette, nucleus and centrosome. During fertilization, UBR7 remained with the ZP-bound acrosomal shroud following acrosomal exocytosis. Thus, UBR7 is present in the acrosomal cap of round spermatids and within the acrosomal matrix of mature boar spermatozoa. These data provide the first evidence of ubiquitin ligase activity in mammalian spermatozoa and indicate UBR7 involvement in spermiogenesis. |
| Databáze: | OpenAIRE |
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