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PMID: 2987274 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

The small subunit of ribonucleotide reductase is encoded by one of the most abundant translationally regulated maternal RNAs in clam and sea urchin eggs.

The Journal of cell biology ·Vol. 100 ·No. 6 ·1985-06-00 ·Pages 1968-76

Standart NM, Bray SJ, George EL, Hunt T, Ruderman JV

Abstract

In both clam oocytes and sea urchin eggs, fertilization triggers the synthesis of a set of proteins specified by stored maternal mRNAs. One of the most abundant of these (p41) has a molecular weight of 41,000. This paper describes the identification of p41 as the small subunit of ribonucleotide reductase, the enzyme that provides the precursors necessary for DNA synthesis. This identification is based mainly on the amino acid sequence deduced from cDNA clones corresponding to p41, which shows homology with a gene in Herpes Simplex virus that is thought to encode the small subunit of viral ribonucleotide reductase. Comparison with the B2 (small) subunit of Escherichia coli ribonucleotide reductase also shows striking homology in certain conserved regions of the molecule. However, our attention was originally drawn to protein p41 because it was specifically retained by an affinity column bearing the monoclonal antibody YL 1/2, which reacts with alpha-tubulin (Kilmartin, J. V., B. Wright, and C. Milstein, 1982, J. Cell Biol., 93:576-582). The finding that this antibody inhibits the activity of sea urchin embryo ribonucleotide reductase confirmed the identity of p41 as the small subunit. The unexpected binding of the small subunit of ribonucleotide reductase can be accounted for by its carboxy-terminal sequence, which matches the specificity requirements of YL 1/2 as determined by Wehland et al. (Wehland, J., H. C. Schroeder, and K. Weber, 1984, EMBO [Eur. Mol. Biol. Organ.] J., 3:1295-1300). Unlike the small subunit, there is no sign of synthesis of a corresponding large subunit of ribonucleotide reductase after fertilization. Since most enzymes of this type require two subunits for activity, we suspect that the unfertilized oocytes contain a stockpile of large subunits ready for combination with newly made small subunits. Thus, synthesis of the small subunit of ribonucleotide reductase represents a very clear example of the developmental regulation of enzyme activity by control of gene expression at the level of translation.

MeSH Terms
Amino Acid Sequence Animals Antibodies, Monoclonal Base Sequence Bivalvia Chromatography, Affinity DNA Escherichia coli/genetics Female Fertilization Molecular Weight Oocytes/enzymology Ovum/enzymology Protein Biosynthesis RNA, Messenger/genetics Ribonucleotide Reductases/genetics Sea Urchins Simplexvirus/genetics Tubulin/immunology
Chemicals
Antibodies, Monoclonal RNA, Messenger Tubulin DNA Ribonucleotide Reductases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Standart N M
Bray S J
George E L
Hunt T
Ruderman J V
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48 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1985-06-00
Pages
1968-76
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2113603
Subset
IM
Grants
NIGMS NIH HHS · GM-31136-05 · United States
Databases
GENBANK
M23795
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