title: Basic nuclear proteins of the histone-less eukaryote Crypthecodinium cohnii (Pyrrhophyta): two-dimensional electrophoresis and DNA-binding properties.
abstract: Unlike typical eukaryotes, the Dinoflagellate Crypthecodinium cohnii does not contain histones but six major basic, low molecular weight nuclear proteins which represent only 10% of the DNA mass and differ from histones in their electrophoretic and DNA-binding properties. These proteins are resolved in two-dimensional electrophoresis (AUT-PAGE x SDS-PAGE). Three proteins with an apparent molecular mass of 16, 16.5 and 17 kDa (p16, p16.5 and p17) are present in addition to the major 14 kDa basic nuclear component (HCc). HCc itself is resolved in three proteins (alpha, beta and gamma). When the proteins are not reduced with 2-mercaptoethanol before 2D-PAGE, the migration of HCc alpha, beta and gamma is modified in a way which suggests the formation of both inter- and intramolecular disulfide bridges and thus, the presence of at least two cysteines. The amino-acid analysis of HCc proteins resolved in 2D gels confirms that they are lysine-rich. HCc alpha, beta and gamma as well as p16, p16.5 and p17 are removed from isolated chromatin with 0.6 M NaCl, indicating that their affinity for DNA in vivo is lower than that of core histones. Furthermore, in vitro, they bind more tightly to single-stranded than to double-stranded DNA.
authors.affiliation: Biozentrum, Basel, Switzerland.
authors.forename: G
authors.initials: G
authors.lastname: Vernet
authors.forename: M
authors.initials: M
authors.lastname: Sala-Rovira
authors.forename: M
authors.initials: M
authors.lastname: Maeder
authors.forename: F
authors.initials: F
authors.lastname: Jacques
authors.forename: M
authors.initials: M
authors.lastname: Herzog
mesh_terms: D000818:Animals; D002417:Cattle; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D004141:Dinoflagellida / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000302:isolation & purification; D015151:Immunoblotting; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000302:isolation & purification* / Q000378:metabolism; D013950:Thymus Gland / Q000032:analysis
chemical_list: D004268:DNA-Binding Proteins; D006657:Histones; D009687:Nuclear Proteins