Breakdown of cytoskeletal proteins during meiosis of starfish oocytes and proteolysis induced by calpain.
Meiosis reinitiation in starfish oocytes is characterized by Ca(2+) transients in the cytosol and in the nucleus and is accompanied by the disassembly of the nuclear envelope, a process which is likely to be mediated by the cleavage of selected proteins. We have used mass spectrometry analysis (mass profile fingerprinting) on 2D polyacrylamide gels of extracts of oocytes in which meiosis resumption was induced by 1-methyladenine and have identified five proteins that were specifically degraded: alpha-tubulin, lamin B, dynamin, and two kinds of actin. They are all components of the cytoskeleton or associated with it. We then investigated whether calpain, which is activated by the increase in cell Ca(2+), could cleave the same proteins that became degraded under the influence of 1-methyladenine and thus be involved in nuclear membrane breakdown. The investigation was prompted by the finding that microinjection of calpain into the nuclei of prophase arrested oocytes induced meiosis in the absence of 1-methyladenine. Incubation of prophase arrested (disrupted) oocytes with calpain produced a 2D gel protein pattern in which some of the degradation products coincided with those seen in oocytes challenged with 1-methyladenine.
Santella
L
L
Laboratory of Cell Biology, Stazione Zoologica "A. Dohrn", Napoli, I-80121, Italy. santella@alpha.szn.it
Kyozuka
K
K
Hoving
S
S
Munchbach
M
M
Quadroni
M
M
Dainese
P
P
Zamparelli
C
C
James
P
P
Carafoli
E
E
D000199:Actins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000378:metabolism* / Q000494:pharmacology*; D003599:Cytoskeleton / Q000378:metabolism*; D034281:Dynamins; D015180:Electrophoresis, Gel, Two-Dimensional; D020558:GTP Phosphohydrolases / Q000032:analysis / Q000378:metabolism; D034921:Lamin Type B; D034882:Lamins; D008540:Meiosis / Q000187:drug effects / Q000502:physiology*; D008845:Microinjections; D009685:Nuclear Envelope / Q000187:drug effects / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000201:enzymology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013215:Starfish; D013379:Substrate Specificity / Q000502:physiology; D014404:Tubulin / Q000032:analysis / Q000378:metabolism
D000199:Actins; D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; D014404:Tubulin; D002154:Calpain; D020558:GTP Phosphohydrolases; D034281:Dynamins; D002118:Calcium