Molecular cloning and immunolocalization of two variants of the major basic nuclear protein (HCc) from the histone-less eukaryote Crypthecodinium cohnii (Pyrrhophyta).
Two clones that encode variants (HCc1 and HCc2) of the major basic nuclear protein of the dinoflagellate Crypthecodinium cohnii, were identified by immunoscreening of a cDNA expression library. The first clone carries a full-length cDNA with an open reading frame (HCc1) encoding 113 amino acids. The cDNA from the second clone lacks some of the 5' end, and the coding sequence is only 102 residues. The two proteins display 77% sequence similarity and their NH2-ends are homologous to the NH2-peptide of the HCc protein determined by P. Rizzo. The amino acid composition, which confirms the basic nature of lysine-rich HCc proteins, differs markedly from other known DNA-binding proteins such as histones, HMGs or prokaryotic histone-like proteins. No convincing homology was found with other proteins. HCc antigens were localized on C. cohnii by immunofluorescence, and by electron microscopy (EM) with immunogold labelling. HCc proteins are mainly detected at the periphery of the permanently condensed chromosomes, where active chromatin is located, as well as in the nucleolar organizing region (NOR). This suggests that these basic, non-histone proteins, with a moderate affinity for DNA, are involved at some level in the regulation of gene expression.
Sala-Rovira
M
M
Département de Biologie Cellulaire et Moléculaire, Université de Paris, VI CNRS UA 117, Banyuls-sur-Mer, France.
Geraud
M L
ML
Caput
D
D
Jacques
F
F
Soyer-Gobillard
M O
MO
Vernet
G
G
Herzog
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004247:DNA; D004268:DNA-Binding Proteins; D004141:Dinoflagellida / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000302:isolation & purification; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000235:genetics* / Q000302:isolation & purification; D016415:Sequence Alignment
D004268:DNA-Binding Proteins; C072291:HCc protein, Crypthecodinium cohnii; D009687:Nuclear Proteins; D015800:Protozoan Proteins; D004247:DNA