DNA (cytosine-5) methyltransferase turnover and cellular localization in developing Paracentrotus lividus sea urchin embryo.
The turnover and localization of the enzyme DNA (cytosine-5) methyltransferase (Dnmt1) were studied during Paracentrotus lividus sea urchin embryo development using antibody preparations against the NH(2) and COOH-terminal regions of the molecule. The antibodies reveal, by Western blots and whole-mount analyses, that the enzyme is differently required during embryonic development. The changeover point is at blastula stage, where a proteolytic mechanism hydrolyses the enzyme present in all embryonic cells by removing a peptide of about 45 kDa from the amino terminal region of the 190 kDa enzyme initially synthesized on maternal transcripts. The resulting 145 kDa enzyme shows modified catalytic properties, different antibody reactivity and is rapidly destroyed in the few hours before gastrulation. At more advanced stages of development the enzyme is newly synthesized but only in particular cell types, among which neurons. The data show that Dnmt1 is removed from embryonic cells before gastrulation to be synthesized again at different levels in different cell types, indicating that the concentration of Dnmt1 is critical for the various differentiated cells of the developing sea urchin embryo.
Di Giaimo
R
R
Department of Genetics, General and Molecular Biology, University of Naples Federico II, Via Mezzocannone 8, 80134 Naples, Italy.
Locascio
A
A
Aniello
F
F
Branno
M
M
del Gaudio
R
R
Potenza
N
N
Geraci
G
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000201:enzymology*; D047108:Embryonic Development; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D011233:Precipitin Tests; D012617:Sea Urchins / Q000201:enzymology*; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D004247:DNA; D004248:DNA (Cytosine-5-)-Methyltransferases