An assessment of the masked message hypothesis: Sea urchin egg messenger ribonucleoprotein complexes are efficient templates for in vitro protein synthesis

Randall T. Moon, Michael Danilchik, Merrill B. Hille

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The rate of protein synthesis in unfertilized sea urchin eggs is very low, although all components for protein synthesis are present. To determine whether egg messenger RNAs are unavailable for translation because of "masking" by phenol-soluble inhibitors, crude and purified nonpolyribosomal messenger ribonucleoprotein complexes (mRNPs) from eggs of Strongylocentrotus purpuratus were translated in vitro in a wheat germ cell-free system. Crude and purified egg mRNPs were nearly as translatable as the mRNAs extracted from the mRNPs, suggesting that the mRNPs were not masked. No difference in the relative translational activities of mRNPs and their constituent mRNAs was revealed by isolating the mRNPs in buffers of different ionic strength or in the presence of protease and ribonuclease inhibitors. Furthermore, kinetic analysis of the in vitro translation and translation of the mRNPs and mRNAs at several concentrations of K+, Mg2+, and template all indicate that mRNPs are efficient templates for directing protein synthesis. Separation on polyacrylamide gels of the products of in vitro and in vivo translation demonstrated that both mRNPs and mRNAs extracted from the mRNPs synthesized in vitro high-molecular-weight polypeptides, some of which were also synthesized in vivo. Although sea urchin egg mRNPs may not be masked, there are several alternative mechanisms for regulating translation in the egg.

Original languageEnglish (US)
Pages (from-to)389-403
Number of pages15
JournalDevelopmental Biology
Volume93
Issue number2
DOIs
StatePublished - 1982
Externally publishedYes

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Sea Urchins
Ovum
Proteins
Messenger RNA
Eggs
messenger ribonucleoprotein
In Vitro Techniques
Strongylocentrotus purpuratus
Cell-Free System
Ribonucleases
Phenol
Protease Inhibitors
Germ Cells
Osmolar Concentration
Triticum
Buffers
Molecular Weight

ASJC Scopus subject areas

  • Developmental Biology

Cite this

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title = "An assessment of the masked message hypothesis: Sea urchin egg messenger ribonucleoprotein complexes are efficient templates for in vitro protein synthesis",
abstract = "The rate of protein synthesis in unfertilized sea urchin eggs is very low, although all components for protein synthesis are present. To determine whether egg messenger RNAs are unavailable for translation because of {"}masking{"} by phenol-soluble inhibitors, crude and purified nonpolyribosomal messenger ribonucleoprotein complexes (mRNPs) from eggs of Strongylocentrotus purpuratus were translated in vitro in a wheat germ cell-free system. Crude and purified egg mRNPs were nearly as translatable as the mRNAs extracted from the mRNPs, suggesting that the mRNPs were not masked. No difference in the relative translational activities of mRNPs and their constituent mRNAs was revealed by isolating the mRNPs in buffers of different ionic strength or in the presence of protease and ribonuclease inhibitors. Furthermore, kinetic analysis of the in vitro translation and translation of the mRNPs and mRNAs at several concentrations of K+, Mg2+, and template all indicate that mRNPs are efficient templates for directing protein synthesis. Separation on polyacrylamide gels of the products of in vitro and in vivo translation demonstrated that both mRNPs and mRNAs extracted from the mRNPs synthesized in vitro high-molecular-weight polypeptides, some of which were also synthesized in vivo. Although sea urchin egg mRNPs may not be masked, there are several alternative mechanisms for regulating translation in the egg.",
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