In vitro decondensation of the sperm chromatin in Holothuria tubulosa (sea cucumber) not affecting proteolysis of basic nuclear proteins.
Sea urchin and sea star oocyte extracts contain proteolytic activities that are active against sperm basic nuclear proteins (SNBP). This SNBP degradation has been related to the decondensation of sperm chromatin as a possible model to male pronuclei formation. We have studied the presence of this proteolytic activity in Holothuria tubulosa (sea cucumber) and its possible relationship with sperm nuclei decondensation. The mature oocyte extracts from H. tubulosa contain a proteolytic activity to SNBP located in the macromolecular fraction of the egg-jelly layer. SNBP degradation occurred both on sperm nuclei and on purified SNBP, histones being more easily degraded than protein Ø(o) (sperm-specific protein). SNBP degradation was found to be dependent on concentration, incubation time, presence of Ca(2+), pH, and this activity could be a serine-proteinase. Thermal denaturalization of the oocyte extracts (80 degrees C, 10-15 min) inactivates its proteolytic activity on SNBP but does not affect sperm nuclei decondensation. These results would suggest that sperm nuclei decondensation occurs by a mechanism different from SNBP degradation. Thus, the sperm nuclei decondensation occurs by a thermostable factor(s) and the removal of linker SNBP (H1 and protein Ø(o)) will be a first condition in the process of sperm chromatin remodeling.
del Valle
Luis J
LJ
Center de Biotecnología Molecular, Departament d'Enginyeria Química, EUETIB, Consorci Escola Industrial de Barcelona, Universitat Politècnica de Catalunya, 08036 Barcelona, Spain. luis.javier.del.valler@upc.edu
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002843:Chromatin / Q000378:metabolism*; D005260:Female; D006863:Hydrogen-Ion Concentration; D016014:Linear Models; D008297:Male; D008858:Microscopy, Phase-Contrast; D009687:Nuclear Proteins / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D012616:Sea Cucumbers / Q000378:metabolism*; D013094:Spermatozoa / Q000166:cytology* / Q000648:ultrastructure
D002843:Chromatin; D009687:Nuclear Proteins; C026588:sperm basic nuclear proteins; D002118:Calcium