Protein deubiquitination during oocyte maturation influences sperm function during fertilisation, antipolyspermy defense and embryo development.
Ubiquitination is a covalent post-translational modification of proteins by the chaperone protein ubiquitin. Upon docking to the 26S proteasome, ubiquitin is released from the substrate protein by deubiquitinating enzymes (DUBs). We hypothesised that specific inhibitors of two closely related oocyte DUBs, namely inhibitors of the ubiquitin C-terminal hydrolases (UCH) UCHL1 (L1 inhibitor) and UCHL3 (L3 inhibitor), would alter porcine oocyte maturation and influence sperm function and embryo development. Aberrant cortical granule (CG) migration and meiotic spindle defects were observed in oocytes matured with the L1 or L3 inhibitor. Embryo development was delayed or blocked in oocytes matured with the general DUB inhibitor PR-619. Aggresomes, the cellular stress-inducible aggregates of ubiquitinated proteins, formed in oocytes matured with L1 inhibitor or PR-619, a likely consequence of impaired protein turnover. Proteomic analysis identified the major vault protein (MVP) as the most prominent protein accumulated in oocytes matured with PR-619, suggesting that the inhibition of deubiquitination altered the turnover of MVP. The mitophagy/autophagy of sperm-contributed mitochondria inside the fertilised oocytes was hindered by DUB inhibitors. It is concluded that DUB inhibitors alter porcine oocyte maturation, fertilisation and preimplantation embryo development. By regulating the turnover of oocyte proteins and mono-ubiquitin regeneration, the DUBs may promote the acquisition of developmental competence during oocyte maturation.
Division of Animal Sciences, University of Missouri, S141 ASRC, 920 East Campus Drive, Columbia, MO65211-5300, USA.
Young-Joo
YJ
Yi
Division of Animal Sciences, University of Missouri, S141 ASRC, 920 East Campus Drive, Columbia, MO65211-5300, USA.
Miriam
M
Sutovsky
Division of Animal Sciences, University of Missouri, S141 ASRC, 920 East Campus Drive, Columbia, MO65211-5300, USA.
Won-Hee
WH
Song
Division of Animal Sciences, University of Missouri, S141 ASRC, 920 East Campus Drive, Columbia, MO65211-5300, USA.
Peter
P
Sutovsky
D000818:Animals; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005306:Fertilization / Q000187:drug effects / Q000502:physiology*; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D011247:Pregnancy; D040901:Proteomics; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013552:Swine; D043222:Ubiquitin Thiolesterase / Q000037:antagonists & inhibitors*
D015853:Cysteine Proteinase Inhibitors; D043222:Ubiquitin Thiolesterase