Subcellular localization of nardilysin during mouse oocyte maturation.
We have previously shown that the peptidase, nardilysin, contains a bipartite nuclear localization signal that permits the enzyme to cycle between the nucleus and cytoplasm. In the present study, we report that nardilysin accumulates in the nucleus of an oocyte as a function of its maturation. Nardilysin is predominantly localized in the cytoplasm of an oocyte when initially placed into culture. The enzyme starts to accumulate in the nucleus within 30 min of in vitro culture. After 3 h, nardilysin is found as a spherical structure surrounded by condensed chromosomal DNA. After 18 h of in vitro culture, it co-localizes with beta-tubulin at the spindle apparatus. Cilostamide, a phosphodiesterase 3A inhibitor that inhibits meiosis, blocks accumulation of nuclear nardilysin. This finding demonstrates that the nuclear entry of nardilysin is tightly controlled in the oocyte. Taken together, these experiments strongly suggest a role for nardilysin in meiosis through its dynamic translocation from cytosol to nucleus, and then to the spindle apparatus.
Ma
Zhangliang
Z
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky Medical Center, 800 Rose St., Lexington, KY 40536, USA. maz@uky.edu
Wang
Xuebing
X
Hockman
Steven
S
Snow
E Charles
EC
Hersh
Louis B
LB
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000818:Animals; D005260:Female; D066298:In Vitro Techniques; D008540:Meiosis; D008666:Metalloendopeptidases / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D015363:Quinolones / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D014404:Tubulin / Q000378:metabolism
D015363:Quinolones; D014404:Tubulin; C021294:cilostamide; D008666:Metalloendopeptidases; C087976:nardilysin