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 Nepsilon(gamma-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 internucleosomal. When sperm nuclei were incubated with monodansylcadaverine, a fluorescent amine, in the presence or absence of Ca(2+), histone H2B was modified only in the presence of Ca(2+). 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 Ca(2+)-dependent nuclear transglutaminase. Treatment of sperm with the egg jelly, which activates Ca(2+) 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.
Nunomura
Kazuto
K
Department of Applied Biochemistry, Hiroshima University, Higashi-hiroshima, Hiroshima, Japan.
Kawakami
Satoru
S
Shimizu
Takahiko
T
Hara
Tomohiro
T
Nakamura
Kazuhiro
K
Terakawa
Yudai
Y
Yamasaki
Akiko
A
Ikegami
Susumu
S
D020101:Acrosome Reaction / Q000187:drug effects / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002103:Cadaverine / Q000031:analogs & derivatives; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology*; D002843:Chromatin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000494:pharmacology; D019281:Dimerization; D005260:Female; D006657:Histones / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008836:Micrococcal Nuclease / Q000378:metabolism; D008969:Molecular Sequence Data; D009707:Nucleosomes / Q000378:metabolism*; D010063:Ovum / Q000737:chemistry / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013154:Spleen / Q000166:cytology; D013215:Starfish / Q000502:physiology*; D011503:Transglutaminases / Q000378:metabolism*
D002843:Chromatin; D003432:Cross-Linking Reagents; D006657:Histones; D009707:Nucleosomes; D011503:Transglutaminases; D008836:Micrococcal Nuclease; D002103:Cadaverine; D002118:Calcium