TAILIEUCHUNG - Báo cáo khoa học: In vivo cross-linking of nucleosomal histones catalyzed by nuclear transglutaminase in starfish sperm and its induction by egg jelly triggering the acrosome reaction

A histone heterodimer, designated as p28, which contains an N e (c-glutamyl)lysine cross-link between Gln9 of histone H2B and Lys5 or Lys12 of histone H4, is present in starfish (Asterina pectinifera) sperm. Treatment of spermnuclei with micrococcal nuclease produced soluble chromatin, which was size-fractionated by sucrose-gradient centrifugation to give p28-containing oligonucleosome and p28-free mono-nucleosome fractions, indicating that the cross-link is inter-nucleosomal. | Eur. J. Biochem. 270 3750-3759 2003 FEBS 2003 doi In vivo cross-linking of nucleosomal histones catalyzed by nuclear transglutaminase in starfish sperm and its induction by egg jelly triggering the acrosome reaction Kazuto Nunomura Satoru Kawakamit Takahiko Shimizut Tomohiro Hara Kazuhiro Nakamura Yudai Terakawa Akiko Yamasaki and Susumu Ikegamiị Department of Applied Biochemistry Hiroshima University Higashi-hiroshima Hiroshima Japan A histone heterodimer designated as p28 which contains an NE y-glutamyl lysine cross-link between Gln9 of histone H2B and Lys5 or Lys12 of histone H4 is present in starfish Asterina pectinifera sperm. Treatment of sperm nuclei with micrococcal nuclease produced soluble chromatin which was size-fractionated by sucrose-gradient centrifugation to give p28-containing oligonucleosome and p28-free mononucleosome fractions indicating that the cross-link is inter-nucleosomal. When sperm nuclei were incubated with monodansylcadaverine a fluorescent amine in the presence or absence of Ca2 histone H2B was modified only in the presence of Ca2 . Gln9 in the N-terminal region was modified but the other Gln residues located in the internal region were not suggesting that the modification takes place on the surface of the nucleosome core by the in situ action of a Ca2 -dependent nuclear transglutaminase. Treatment of sperm with the egg jelly which activates Ca2 influx to induce the acrosome reaction resulted in a significant elevation of the p28 content in the nucleus. This is the first demonstration of an in vivo activation of transglutaminase leading to the formation of a cross-link in intracellular proteins. Keywords starfish sperm transglutaminase chromatin acrosome reaction. The nucleosome which contains four core histones H2A H2B H3 and H4 is the primary unit of chromatin. The higher-order chromatin structure is stabilized by inter-nucleosomal interactions involving the N-terminal region of core histones and by

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