Dissociation of protamine-DNA complexes by Xenopus nucleoplasmin and minichromosome assembly in vitro.
Nucleoplasmin, an acidic thermostable protein abundant in the nucleus of Xenopus laevis oocytes, has been found to dissociate complexes of pUC19 DNA and protein phi 1, an intermediate protamine present in ripe sperm from the mollusc Mytilus edulis. Cruder preparations of nucleoplasmin, such as the amphibian oocyte S150 extract and its thermostable fraction, also dissociate the heterologous DNA-phi 1 complexes and, in addition, promote the assembly of plasmid DNA into a minichromosome displaying regular nucleosomal periodicity, as revealed by micrococcal nuclease digestion. In contrast, purified nucleoplasmin complemented with rat hepatocyte core histone octamers in the presence of DNA topoisomerase I, although capable of inducing nucleoprotein formation onto the complexed DNA, fails to position nucleosomes at the native spacings seen in chromatin in vivo. These data favour the existence of a general mechanism to bring about, in a concerted manner, removal of sperm-specific nuclear proteins and reconstitution of somatic chromatin following fertilization.
Ruiz-Lara
S A
SA
Departamento de Biología Molecular y Celular, Centro de Investigación y Desarrollo del CSIC, Barcelona, Spain.
Cornudella
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L
Rodríguez-Campos
A
A
D000818:Animals; D001667:Binding, Competitive; D049872:Bivalvia; D002499:Centrifugation, Density Gradient; D002875:Chromosomes / Q000502:physiology; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins; D005260:Female; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D009687:Nuclear Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D056489:Nucleoplasmins; D009865:Oocytes / Q000502:physiology*; D010750:Phosphoproteins; D011479:Protamines / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D013094:Spermatozoa / Q000502:physiology*; D014982:Xenopus laevis
D004268:DNA-Binding Proteins; D006657:Histones; D009687:Nuclear Proteins; D056489:Nucleoplasmins; D010750:Phosphoproteins; D011479:Protamines; C082114:phi1 protein, Mytilus edulis; D004247:DNA