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Identification of a cysteine protease responsible for degradation of sperm histones during male pronucleus remodeling in sea urchins
Author(s) -
Imschenetzky Maria,
Díaz Fredy,
Montecino Martin,
Sierra Felipe,
Puchi Marcia
Publication year - 1997
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/(sici)1097-4644(19971201)67:3<304::aid-jcb3>3.0.co;2-#
Subject(s) - pepstatin , leupeptin , protease , biochemistry , cysteine protease , histone , male pronucleus , biology , sperm , pmsf , serine protease , proteases , protamine , chromatin remodeling , chemistry , microbiology and biotechnology , enzyme , genetics , embryogenesis , heparin , zygote , dna , gene
We have identified a 60‐kDa cysteine protease that is associated with chromatin in sea urchin zygotes. This enzyme was found to be present as a proenzyme in unfertilized eggs and was activated shortly after fertilization. At a pH of 7.8–8.0, found after fertilization, the enzyme degraded the five sperm‐specific histones (SpH), while the native cleavage‐stage (CS) histone variants remained unaffected. Based on its requirements for reducing agents, its inhibition by sulfhydryl blocking compounds and its sensitivity to the cysteine‐type protease inhibitors (2S,3S)‐ translator ‐epoxysuccinyl‐L‐leucyl‐amido‐3‐methylbutane‐ethyl‐ester (E‐64 d), cystatin and leupeptin, this protease can be defined as a cysteine protease. Consistently, this protease was not affected by the serine‐type protease inhibitors phenylmethylsulfonyl fluoride (PMSF) and pepstatin. The substrate selectivity and pH modulation of the protease activity strongly suggest its role in the removal of sperm‐specific histones, which determines sperm chromatin remodeling after fertilization. This suggestion was further substantiated by the inhibition of sperm histones degradation in vivo by E‐64 d. Based on these three lines of evidence, we postulate that this cysteine protease is responsible for the degradation of sperm‐specific histones which occurs during male pronucleus formation. J. Cell. Biochem. 67:304–315, 1997. © 1997 Wiley‐Liss, Inc.