Cyclic expression of a nuclear protein in a dinoflagellate.
Nuclei of the dinoflagellate Crypthecodinium cohnii strain Whd were isolated and nuclear proteins were extracted in three fractions, corresponding to the increasing affinity of these proteins to genomic DNA. One fraction contained two major bands (48- and 46-kDa) and antibodies specific to this fraction revealed two major bands by Western blot on nuclear extracts, corresponding to the 46- and 48-kDa bands. The 48-kDa protein was detected in G1 phase but not in M phase cells. An expression cDNA library of C. cohnii was screened with these antibodies, and two different open reading frames were isolated. Dinoflagellate nuclear associated protein (Dinap1), one of these coding sequences, was produced in E. coli and appeared to correspond to the 48-kDa nuclear protein. No homologue of this sequence was found in the data bases, but two regions were identified, one including two putative zinc finger repeats, and one coding for two potential W/W domains. The second coding sequence showed a low similarity to non-specific sterol carrier proteins. Immunocytolocalization with specific polyclonal antibodies to recombinant Dinap1 showed that the nucleus was immunoreactive only during the G1 phase: the nucleoplasm was immunostained, while chromosome cores and nuclear envelopes were negative.
Bhaud
Y
Y
Observatoire Océanologique de Banyuls, Laboratoire Arago, UMR CNRS 7628, Banyuls-sur-Mer France.
Géraud
M L
ML
Ausseil
J
J
Soyer-Gobillard
M O
MO
Moreau
H
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D016054:DNA, Protozoan / Q000032:analysis; D004141:Dinoflagellida / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D051034:Inhibitor of Apoptosis Proteins; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011817:Rabbits; D017422:Sequence Analysis, DNA; D016335:Zinc Fingers
C111891:DIAP1 protein, Drosophila; D016054:DNA, Protozoan; D029721:Drosophila Proteins; D051034:Inhibitor of Apoptosis Proteins; D019476:Insect Proteins; D009687:Nuclear Proteins; D015800:Protozoan Proteins