A characterization of cytostatic factor activity from Xenopus eggs and c-mos-transformed cells.
In Xenopus oocytes, the mos proto-oncogene product is required during meiosis I for the activation of maturation promoting factor (MPF) and the subsequent breakdown of the germinal vesicle (GVBD). In addition, the mos product has been shown to be a candidate "initiator" of meiotic maturation and is an active component of cytostatic factor (CSF), an activity responsible for metaphase II arrest. Here we demonstrate that pp39mos is required throughout oocyte maturation. We found that in progesterone stimulated oocytes, depletion of mos RNA immediately before GVBD terminally decreased MPF. Likewise, oocytes depleted of mos RNA and induced to mature with crude MPF proceeded through GVBD but lacked the MPF activity required to arrest mature oocytes at metaphase II. Thus, during maturation the mos product is required, directly or indirectly, to sustain MPF activity. On the other hand, mouse NIH/3T3 cells transformed by the constitutive expression of pp39mosxc possessed CSF activity but lacked constitutive levels of MPF or its associated histone H1 kinase activity. Moreover, cytosols prepared from transformed NIH/3T3 cells or Xenopus eggs had similar levels of CSF activity, but pp39mos levels were greater than 40-fold higher in the transformed cell extract. These analyses show that maintenance of CSF during interphase does not result in the maintenance of MPF.
Daar
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ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 21702.
Paules
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Vande Woude
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GF
D000818:Animals; D000970:Antineoplastic Agents / Q000032:analysis / Q000378:metabolism*; D002461:Cell Line, Transformed; D005260:Female; D005347:Fibroblasts / Q000737:chemistry / Q000378:metabolism; D016200:Maturation-Promoting Factor / Q000032:analysis / Q000378:metabolism; D000090204:Mesothelin; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009866:Oogenesis; D010063:Ovum / Q000737:chemistry / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000378:metabolism*; D017231:Proto-Oncogene Proteins c-mos; D014981:Xenopus / Q000378:metabolism*
D000970:Antineoplastic Agents; C000715528:Msln protein, mouse; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D017231:Proto-Oncogene Proteins c-mos; D016200:Maturation-Promoting Factor; D000090204:Mesothelin