Abstract
Cell-free extracts of Arbacia eggs (Arbacia punctulata) apparently do not contain an enzymatic system for the reduction of ribonucleotides to deoxyribonucleotides. However, during an interval of 5 hr after fertilization at 23 degrees , an enzymatic system is produced that is capable of catalyzing the reduction of CDP to dCDP in the presence of Mg(2+), ethylenediaminetetraacetate, ATP, and a reducing agent, dithiothreitol. The activity is first seen about 1 hr after fertilization, and reaches a peak at about 5 hr. The appearance of the ribonucleotide reductase is prevented by the addition of emetine or puromycin, inhibitors of protein synthesis, to the cells before fertilization. Inclusion of actinomycin D in the cell suspension at a concentration sufficient to inhibit synthesis of messenger RNA does not appreciably affect the production of the enzyme activity. Preexisting, maternal RNA is thus used for synthesis of reductase. Ribonucleotide reductase may, therefore, represent the first example of an enzyme system absent in unfertilized eggs that is produced in response to fertilization.
MeSH Terms
Adenosine Triphosphate
Alcohol Oxidoreductases/biosynthesis
Animals
Cell Fractionation
Cell-Free System
Centrifugation, Density Gradient
Chromatography, Ion Exchange
Cytosine Nucleotides/metabolism
Dactinomycin/pharmacology
Deoxyribonucleotides/biosynthesis
Dithiothreitol
Edetic Acid
Embryo, Nonmammalian/enzymology
Emetine/pharmacology
Enzyme Induction
Female
Fertilization
Magnesium
Nucleotides/biosynthesis
Ovum/drug effects
Proteins/antagonists & inhibitors
Puromycin/pharmacology
RNA, Messenger/antagonists & inhibitors
Ribonucleotides
Sea Urchins/embryology
Chemicals
Cytosine Nucleotides
Deoxyribonucleotides
Nucleotides
Proteins
RNA, Messenger
Ribonucleotides
Dactinomycin
Puromycin
Adenosine Triphosphate
Edetic Acid
Alcohol Oxidoreductases
Magnesium
Dithiothreitol
Emetine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Noronha J M
Sheys G H
Buchanan J M
References (22)
22 references, click to expand
-
Formation of deoxycytidine phosphates from cytidine phosphates in extracts from Escherichia coli.
J Biol Chem. 1961 Apr;236:1150-7
PMID: 13740426
-
The enzymatic deamination of 6-aminopyrimidine deoxyribonucleotides. I. The enzymatic deamination of deoxycytidine 5'-phosphate and of 5-methyldeoxycytidine 5-methyldeoxycytidine 5'-phosphate.
J Biol Chem. 1960 Mar;235:706-13
PMID: 14442222
-
Effects of actinomycin D on macromolecule synthesis and early development in sea urchin eggs.
Biochem Biophys Res Commun. 1963 Feb 18;10:321-6
PMID: 13960718
-
TEMPLATES FOR THE FIRST PROTEINS OF EMBRYONIC DEVELOPMENT.
Proc Natl Acad Sci U S A. 1964 Mar;51:407-14
PMID: 14171452
-
ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEOTIDES. IV. ISOLATION AND CHARACTERIZATION OF THIOREDOXIN, THE HYDROGEN DONOR FROM ESCHERICHIA COLI B.
J Biol Chem. 1964 Oct;239:3436-44
PMID: 14245400
-
ENZYMATIC SYNTHESIS OF DEOXYRIBONUCLEOTIDES. VI. THE CYTIDINE DIPHOSPHATE REDUCTASE SYSTEM FROM NOVIKOFF HEPATOMA.
J Biol Chem. 1964 Oct;239:3453-6
PMID: 14245402
-
Characteristics of the utilization of nucleosides by embryos of Paracentrotus lividus.
J Biol Chem. 1962 Jan;237:143-9
PMID: 14478971
-
Effects of hydroxyurea on development and regeneration.
Nature. 1967 Jun 10;214(5093):1132-3
PMID: 6069549
-
Some effects of deoxyribonucleosides on sea urchin egg development.
Curr Mod Biol. 1968 Jan-Feb;1(5):314-9
PMID: 5689811
-
Thymidine kinase, thymidylate kinase and 32P and [14C] thymidine incorporation into DNA during early embryogenesis of the sea urchin.
Biochim Biophys Acta. 1968 May 21;157(3):546-57
PMID: 5665902
-
A new ribonucleotide reductase system after infection with phage T4.
Proc Natl Acad Sci U S A. 1969 Mar;62(3):829-35
PMID: 4389750
-
Sea urchin nuclear DNA polymerase. II. Changing localization during early development.
Exp Cell Res. 1969 Oct;57(2):305-10
PMID: 5388418
-
Ribonucleotide reductase rom regenerating rat liver.
Eur J Biochem. 1969 Nov;11(1):113-21
PMID: 5353594
-
The relationship of protein synthesis to early transcriptive events in bacteriophage T4-infected Escherichia coli B.
J Biol Chem. 1970 Apr 10;245(7):1575-87
PMID: 4314589
-
Purification and properties of ribonucleotide reductase from leukemic mouse spleen.
J Biol Chem. 1970 Apr 10;245(7):1688-93
PMID: 4314594
-
Purification of a thioredoxin system from yeast.
J Biol Chem. 1970 May 10;245(9):2363-70
PMID: 5442277
-
Cytoplasmic synthesis of RNA in the sea urchin embryo.
Dev Biol. 1970 Jun;22(2):343-65
PMID: 4193236
-
Ribonucleotide reductase and cell proliferation. I. Variations of ribonucleotide reductase activity with tumor growth rate in a series of rat hepatomas.
J Biol Chem. 1970 Oct 25;245(20):5228-33
PMID: 4319235
-
Deoxyribonucleotide pools and deoxyribonucleic acid synthesis in cultured mouse embryo cells.
J Biol Chem. 1970 Oct 25;245(20):5360-8
PMID: 4319240
-
Sea urchin embryos are permeable to actinomycin.
Science. 1971 Feb 19;171(3972):686-9
PMID: 5540310
-
The effect of protein synthesis inhibition on the entry of messenger RNA into the cytoplasm of sea urchin embryos.
J Cell Biol. 1971 Jun;49(3):692-701
PMID: 4997172
-
Oogenetic origin of messenger RNA for embryonic synthesis of microtubule proteins.
Nature. 1972 Jan 28;235(5335):211-4
PMID: 4110479