Peptide coding capacity of polysomal and non-polysomal messenger RNA during growth of animal cells

Lee, Gloria T‐Y.; Engelhardt, Dean

doi:10.1016/0022-2836(79)90278-x

Cited by 45 publications

(15 citation statements)

References 19 publications

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“…A number of laboratories have studied the effect of fresh medium and CHX treatment on the disposition of cellular mRNA (25,44,45). There is general agreement that either treatment leads to an increased loading efficiency of mRNA on ribosomes, resulting in greater polysome size.…”

Section: Discussionmentioning

confidence: 99%

“…In all but a few cases the function and metabolic fate of the stored mRNAs remains unknown. Nonetheless, strong evidence exists that cells select a relatively limited number of mRNAs for inclusion in mRNPs and that these untranslated mRNAs generally have a counterpart in the polysomal mRNA population, with the distribution between these two pools varying widely for individual mRNA species (6,11,20,25).…”

mentioning

confidence: 99%

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Regulation of the utilization of mRNA for eucaryotic elongation factor Tu in Friend erythroleukemia cells.

Rao¹,

Slobin²

1987

Mol. Cell. Biol.

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When Friend erythroleukemia cells were allowed to grow to stationary phase (2 X 10(6) to 3 X 10(6) cells per ml), approximately 60% of the mRNA for eucaryotic elongation factor Tu (eEF-Tu) sedimented at less than or equal to 80S, and most of the remaining factor mRNA was associated with small polysomes. Under the same growth conditions, greater than 90% of the mRNA for eucaryotic initiation factor 4A remained associated with polysomes. The association of eEF-Tu mRNA with polysomes changed dramatically when stationary-phase cells were treated with fresh medium. After 1 h in fresh medium, approximately 90% of eEF-Tu mRNA in Friend cells was found in heavy polysomes. Associated with the shift of eEF-Tu mRNA into heavy polysomes, we found at least a 2.6-fold increase in the synthesis of eEF-Tu in vivo as well as a remarkable 40% decrease in the total amount of eEF-Tu mRNA per cell. Our data raise the possibility that eEF-Tu mRNA that has accumulated in ribonucleoprotein particles in stationary-phase cells is degraded rather than reutilized for eEF-Tu synthesis.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Regulation of the utilization of mRNA for eucaryotic elongation factor Tu in Friend erythroleukemia cells.

Rao¹,

Slobin²

1987

Mol. Cell. Biol.

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“…It should also be pointed out that these proteins do not necessarily represent newly synthesized proteins; they also may be generated by a modification of a second protein. In (25)(26)(27)(28). By using this approach, one also should be able to determine the fate of the mRNAs coding for proteins Q145, Q176, and Q178.…”

Section: Analysis Of Cytoplasmicmentioning

confidence: 99%

Transcriptional and translational control of cytoplasmic proteins after serum stimulation of quiescent Swiss 3T3 cells.

Thomas¹,

Luther²

1981

Proc. Natl. Acad. Sci. U.S.A.

130

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The synthesis ofcytoplasmic proteins from quiescent and serum-stimulated Swiss 3T3 cells was compared by twodimensional polyacrylamide gel electrophoresis. Four new proteins of Mrs 26,000, 28,000, 45,000, and 47,000 designated N26, N28, N45, and N47, which were not detectable in quiescent cells, appeared 60 min after addition ofserum. During the same period, the amount of [asSimethionine incorporated into 10 proteins present in quiescent cells, ranging in Mr from 23,000 to 98,000 and designated Q23-98, increased up to 6-fold, whereas the amount incorporated into three other proteins decreased by a factor of -2. Of the new proteins, N26 was no longer detectable, and the amount of [asS]methionine incorporated into N47 was significantly reduced by 150 min. During this same time, a fifth new protein, N56, appeared, and there was a large increase in the amount of radioactivity incorporated into another protein, Q121. The increases in nine of the proteins were either strongly or completely inhibited by actinomycin D, arguing that the expression of these proteins was under transcriptional control. In contrast, the increases in seven other proteins were unaffected by actinomycin D, suggesting that their expression was under translational control. These proteins will serve as useful markers for determining how cells progress through early lag phase.When quiescent animal cells in culture are stimulated to proliferate by serum or individual growth factors, a number ofcomplex biological processes are activated (1-3). To understand how proliferation is regulated, it will be necessary to determine how each process is controlled, how each is integrated one with another, and how the activation of these processes affects the initiation of DNA synthesis and cell division. Protein synthesis plays a central role. It is required throughout lag phase (time from addition of serum or growth factors until cells enter S phase) for the initiation of DNA synthesis (4), and during S and G2 phases for mitosis and cell division (5). We To understand how the expression of mRNA is regulated during early lag phase, a comprehensive map ofprotein changes after serum stimulation ofquiescent 3T3 cells is required. Next, it must be determined whether these protein changes are controlled by alterations in mRNA expression at the translational or transcriptional level. We describe here a detailed application of two-dimensional polyacrylamide gels (17) to this problem. The results show that, after addition of serum to quiescent 3T3 cells, there are numerous changes in the pattern ofprotein synthesis and that these changes reflect apparent alterations in the expression ofmRNA at both the translational and transcriptional level. MATERIALS AND METHODSCell Culture and Labeling. Swiss mouse 3T3 cells were seeded at 6 X 104 cells per 35-mm tissue culture plate in 2 ml of DME medium (Dulbecco's modified Eagle's medium) containing 10% (vol/vol) fetal calf serum as described (18). After 72 hr the medium was replaced with DME medium S (DME medium supple...

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“…Aliquots (100 pl) of each gradient fraction prepared from Cs2S0, or sucrose gradients (in 0.5 M KCl or SDS/ deoxycholate) were hybridized with [3H]poly(U) to detect poly(A)-containing sequences according to Lee et al [13].…”

Section: Poly ( U ) Hybridizationmentioning

confidence: 99%

A new type of prosome‐like particle, composed of small cytoplasmic RNA and multimers of a 21‐kDa protein, inhibits protein synthesisin vitro

Akhayat

Infante

et al. 1987

European Journal of Biochemistry

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A large fraction of the translationally repressed non-globin messenger RNA in duck erythroblasts is present in non-polyribosomal free mRNP structures which sediment in the 30-40-S range ('35 S'). In 0.5 M KCl, they form core complexes which show a pronounced peak at about 32 S containing mRNA and a discrete spherical RNP particle with a diameter of about 12 nm and the typical morphology of a prosome [H.-P. Schmid et al. (1984) EMBO J . 3,29 -341. Buoyant density measurements and chromatography on oligo(dT)-cellulose indicate that this particle is bound to mRNA; it can be released from the mRNA by treatment of the free mRNP fraction with SDS. This prosome-like particle inhibits the translation of mRNA in vitro. It is composed primarily of multimers of a single 21-kDa protein and at least one species of RNA of about 80-100 nucleotides. It is resistant to dissociation by 2 M Cs2S04 and 1 % SDS; the 21-kDa protein is not attacked by proteinase K unless the particle is extracted with phenol prior to treatment with the protease. The small RNA moiety of the particle hybridizes to the poly(A)-rich mRNA derived from the free mRNPs, as well as to polyribosomal mRNA.These data indicate that prosomes may serve to regulate mRNA translation; they show furthermore that prosome-like particles (about 600 kDa mass) may be built of up to 25 molecules of a single specific protein, rather than of the entire set of about 20 prosomal proteins previously indentified.

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Peptide coding capacity of polysomal and non-polysomal messenger RNA during growth of animal cells

Cited by 45 publications

References 19 publications

Regulation of the utilization of mRNA for eucaryotic elongation factor Tu in Friend erythroleukemia cells.

Regulation of the utilization of mRNA for eucaryotic elongation factor Tu in Friend erythroleukemia cells.

Transcriptional and translational control of cytoplasmic proteins after serum stimulation of quiescent Swiss 3T3 cells.

A new type of prosome‐like particle, composed of small cytoplasmic RNA and multimers of a 21‐kDa protein, inhibits protein synthesisin vitro

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