Purification of myosin mRNP translational control RNA and its inhibition of myosin and globin messenger translation

Kennedy, Doris S.; Siegel, Eric; Heywood, Stuart M.

doi:10.1016/0014-5793(78)80370-6

Cited by 38 publications

(11 citation statements)

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“…These mRNPs as well as mRNAs isolated thereof are inactive as templates in a cell-free translational system, owing to their bound translational inhibitor RNA, which can be dissociated in vitro from poly(A) mRNP or mRNA by poly(U)-Sepharose affinity chromatography in the presence of 10 mM EDTA (2). Similar translational inhibitor RNAs associated with mRNPs have also been detected in the embryonic chicken muscle (3,4), duck erythroblasts (5), rat connective tissue (6) as well as in Artemia salina embryos (7).…”

Section: Introductionmentioning

confidence: 81%

Non-polyadenylated 22 S ribonucleoprotein particle is insensitive to translational inhibitor RNA of cryptobiotic gastrulae of Artemia salina

et al. 1979

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A free cytoplasmic 22 S ribonucleoprotein particle exhibiting a major template activity in rabbit reticulocyte system has been identified in the cryptobiotic gastrulae of Artemia salina. This particle contains non-polyadenylated 9 S messenger RNA which codes primarily for a non-histone basic protein with an apparent molecular weight of 26 000 daltons.We have previously demonstrated the presence of a translational inhibitor RNA which is apparently responsible for transforming polyadenylated messenger ribonucleoproteins into a repressed state in the cryptobiotic gastrulae (Slegers et al., FEBS Letters 80, 390-394, 1977). This inhibitor RNA was found to be completely ineffective on the template activity of non-polyadenylated 22 S messenger ribonucleoprotein, confirming the specificity of this regulatory RNA for polyadenylate sequences.

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Section: Introductionmentioning

confidence: 81%

Non-polyadenylated 22 S ribonucleoprotein particle is insensitive to translational inhibitor RNA of cryptobiotic gastrulae of Artemia salina

et al. 1979

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“…There is suggestive evidence that the myosin mRNA in its free cytoplasmic ribonucleoprotein form is hybridized with a small-M, RNA, the so-called translational control RNA, which prevents its translation [23]. We chose thus an extraction procedure with the intention of leaving intact such putative hybrids [18]. Since, in spite of these precautions, the extracted mRNA was translatable, we conclude that at least in the case of the free cytoplasmic globin mRNA-protein complex, the presence of proteins is required to maintain the mRNA in a repressed conformation.…”

Section: Discussionmentioning

confidence: 99%

“…RNA was recovered by ethanol precipitation. Phenol extraction in the cold was adopted with the aim of maintaining intact possible RNA hybrids [18].…”

Section: Depro Trill Izut Ion ? F ' Rna -Pro Tein Compleses and Pwifimentioning

confidence: 99%

The Translational Repression of Globin mRNA in Free Cytoplasmic Ribonucleoprotein Complexes

Civelli

Víncent

Maundrell

et al. 1980

European Journal of Biochemistry

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The 20-S globin mRNA-protein complex isolated from the post-polyribosomal supernatant of duck reticulocyte lysates was tested for its translational capacities in two different cell-free proteinsynthesizing systems, one derived from wheat germ, the other derived from nuclease-treated rabbit reticulocyte lysates. In both translational systems, the cytoplasmic free globin mRNA-protein complex did not support protein synthesis whereas its fully deproteinized mRNA efficiently directed the synthesis of both the CI and , 4 globin polypeptide chains. Sedimentation of the 20-S mRNAprotein complex through exponential sucrose gradients containing 0.5 M KCl fractionated this complex into four different particles, from which two globin-mRNA-containing core particles were isolated. Neither of these globin-mRNA-containing core particles was translated in either the wheat germ or rabbit reticulocyte lysate system, whereas their extracted mRNA efficiently directed globin synthesis in vitrn.These observations led to the conclusion that factors repressing translational activity are tightly bound to the free cytoplasmic globin mRNA in its ribonucleoprotein form; our data are also consistent with the notion that the associated translational repressing activity is, at least partly, mediated by protein. The results correlate in a functional manner, the different protein compositions observed in the active and inactive forms of globin mRNA-protein complexes isolated respectively from the polyribosomal and the post-polyribosomal cytoplasmic fractions of duck reticulocytes.Although extensive knowledge about the regulation of gene expression in prokaryotes has focused interest on the transcriptional mechanisms of control in eukaryotes, a large body of evidence has accumulated over the years demonstrating the importance of post-transcriptional regulation in animal cells and organisms [l]. The existence of such post-transcriptional controls implies the possibility of differential recognition, and promotion or suppression of already transcribed message in the nucleus or cytoplasm of the cell; necessities and logics of such a system were discussed within the theoretical frame of the cascade regulation hypothesis [ 2 ] . The question arises now about the nature of signals and factors involved in post-transcriptional controls of mRNA formation and expression.

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“…One of these translational control RNAs, tcRNA, has been isolated from eIF-3 preparations of 19-d embryonic chick skeletal muscle (Heywood et al, 1974) and from polysomes and mRNP of 13-14-d embryonic skeletal muscle (Bester et al, 1975;Heywood and Kennedy, 1976). The tcRNA from 19-d embryonic skeletal muscle appears to be the component responsible for message-specific initiation of myoglobin synthesis (see Thompson et al, 1973), whereas the mRNP-associated tcRNA was capable of blocking translation of homologous (skeletal muscle) mRNA (Bester et al, 1975;Heywood et al, 1974Heywood et al, , 1975 by binding the 5' and 3' poly A regions of mRNA (Kennedy et al, 1974(Kennedy et al, , 1978Bester et al, 1975;Heywood et al, 1975).…”

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confidence: 99%

Posttranscriptional control of embryonic rat skeletal muscle protein synthesis. Control at the level of translation by endogenous RNA.

Vanderburg

Nathanson

1988

The Journal of Cell Biology

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Abstract. The onset of muscle cell differentiation is associated with increased transcription of musclespecific mRNA. Studies from this laboratory, using 19-d embryonic rat skeletal muscle, suggest that additional, posttranscriptional controls regulate maturation of muscle tissue via a quantitative effect upon translation, and that the regulatory component may reside within the poly A-RNA pool (Nathanson, M. A., E. W. Bush, and C. Vanderburg. 1986. J. Biol. Chem. 261:1477-1486. To further characterize muscle cell translational control, embryonic and adult total RNA were separated into oligo(dT)cellulose-bound (poly A +) and -unbound (poly A-) pools. Unbound material was subjected to agarose gel electrophoresis to resolve constituents of varying molecular size and mechanically cut into five fractions. Material of each fraction was electroeluted and recovered by precipitation. Equivalent loads of total RNA from 19-20-d embryonic rat skeletal muscle exhibited a 40% translational inhibition in comparison to its adult counterpart. Inhibition was not due to decreased message abundance because embryonic, as well as adult muscle, contained equivalent proportions of poly A ÷ mRNA. An inhibition assay, based upon the translatability of adult RNA and its inhibition by embryonic poly A-RNA, confirmed that inhibition was associated with a 160-2,000-nt poly A-fraction. Studies on the chemical composition of this fraction confirmed its RNA composition, the absence of ribonucleoprotein, and that its activity was absent from similarly fractionated adult RNA. Rescue of inhibition could be accomplished by addition of extra lysate or mRNA; however, smaller proportions of lysate were required, suggesting a strong interaction of inhibitor and components of the translational apparatus. Additional studies demonstrated that the inhibitor acted at the level of initiation, in a dose-dependent fashion. The present studies confirm the existence of translational control in skeletal muscle and suggest that it operates at the embryonic to adult transition. A model of muscle cell differentiation, based upon transcriptional control at the myoblast level, followed by translational regulation at the level of the postmitotic myoblast and/or myotube, is proposed.

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Purification of myosin mRNP translational control RNA and its inhibition of myosin and globin messenger translation

Cited by 38 publications

References 10 publications

Non-polyadenylated 22 S ribonucleoprotein particle is insensitive to translational inhibitor RNA of cryptobiotic gastrulae of Artemia salina

Non-polyadenylated 22 S ribonucleoprotein particle is insensitive to translational inhibitor RNA of cryptobiotic gastrulae of Artemia salina

The Translational Repression of Globin mRNA in Free Cytoplasmic Ribonucleoprotein Complexes

Posttranscriptional control of embryonic rat skeletal muscle protein synthesis. Control at the level of translation by endogenous RNA.

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