scholarly journals Analysis of specific mRNA destabilization during Dictyostelium development

Development ◽  
1989 ◽  
Vol 106 (3) ◽  
pp. 473-481
Author(s):  
G. Mangiarotti ◽  
S. Bulfone ◽  
R. Giorda ◽  
P. Morandini ◽  
A. Ceccarelli ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Dictyostelium Discoideum ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Secondary Effect ◽  
Cell Dissociation ◽  
Specific Mrnas ◽  
Mrna Destabilization ◽  
Selection Of ◽  
Specific Mrna

A number of specific mRNAs are destabilized upon disaggregation of developing Dictyostelium discoideum cells. Analysis of a family of cloned genes indicates that only prespore-enriched mRNAs are affected; constitutive mRNAs that are expressed throughout development and mRNAs that accumulate preferentially in prestalk cells are stable under these conditions. The decay of sensitive prespore mRNAs can be halted by allowing the cells to reaggregate, indicating that destabilization occurs by the progressive selection of individual molecules rather than on all members of an mRNA subpopulation at the time of disaggregation. Individual molecules of the sensitive mRNA species remain engaged in protein synthesis in the disaggregated cells until selected. Destabilization of sensitive mRNAs is induced by cell dissociation even in the presence of concentrations of nogalamycin that inhibit RNA synthesis. The reported prevention of disaggregation-induced mRNA decay by actinomycin D and daunomycin is therefore probably a secondary effect unrelated to the inhibition of transcription.

scholarly journals Specific mRNA destabilization in Dictyostelium discoideum requires RNA synthesis.

1987 ◽  
Vol 7 (12) ◽  
pp. 4585-4588 ◽  
Author(s):  
J F Amara ◽  
H F Lodish
Keyword(s):  
Protein Synthesis ◽  
Dictyostelium Discoideum ◽  
Rna Synthesis ◽  
Protein Synthesis Inhibitors ◽  
Protein And Rna Synthesis ◽  
The Common ◽  
Mrna Destabilization ◽  
Actin Protein ◽  
Specific Mrna

We tested the effects of inhibitors of protein and RNA synthesis on the disaggregation-mediated destabilization of prespore mRNAs in Dictyostelium discoideum. Incubating disaggregated cells with daunomycin to inhibit RNA synthesis prevented the loss of prespore mRNAs, whereas the inhibitor decreased or did not affect levels of the common mRNAs CZ22 and actin. Protein synthesis inhibitors varied in their effects. Cycloheximide, which inhibited protein synthesis almost completely, prevented the loss of the prespore mRNAs, but puromycin, which inhibited protein synthesis less well, did not. These results indicate that the process of specific mRNA destabilization requires the synthesis of RNA and possibly of protein.

Specific mRNA destabilization in Dictyostelium discoideum requires RNA synthesis

1987 ◽  
Vol 7 (12) ◽  
pp. 4585-4588
Author(s):  
J F Amara ◽  
H F Lodish
Keyword(s):  
Protein Synthesis ◽  
Dictyostelium Discoideum ◽  
Rna Synthesis ◽  
Protein Synthesis Inhibitors ◽  
Protein And Rna Synthesis ◽  
The Common ◽  
Mrna Destabilization ◽  
Actin Protein ◽  
Specific Mrna

We tested the effects of inhibitors of protein and RNA synthesis on the disaggregation-mediated destabilization of prespore mRNAs in Dictyostelium discoideum. Incubating disaggregated cells with daunomycin to inhibit RNA synthesis prevented the loss of prespore mRNAs, whereas the inhibitor decreased or did not affect levels of the common mRNAs CZ22 and actin. Protein synthesis inhibitors varied in their effects. Cycloheximide, which inhibited protein synthesis almost completely, prevented the loss of the prespore mRNAs, but puromycin, which inhibited protein synthesis less well, did not. These results indicate that the process of specific mRNA destabilization requires the synthesis of RNA and possibly of protein.

STUDIES ON THE MECHANISM OF ACTION OF ANGIOTENSIN ON FLUID TRANSPORT BY THE MUCOSA OF RAT DISTAL COLON

1972 ◽  
Vol 54 (3) ◽  
pp. 483-492 ◽  
Author(s):  
N. T. DAVIES ◽  
K. A. MUNDAY ◽  
B. J. PARSONS
Keyword(s):  
Protein Synthesis ◽  
Cyclic Amp ◽  
Rna Synthesis ◽  
Fluid Transport ◽  
Actinomycin D ◽  
Distal Colon ◽  
Rat Colon ◽  
Colon Mucosa ◽  
Rat Distal Colon ◽  
Stimulation Of

SUMMARY A study was made of the effects of cyclic AMP, theophylline, cycloheximide, puromycin and actinomycin D on the stimulation by angiotensin of fluid transport by sacs of rat colon mucosa. Cyclic AMP and theophylline, added together or separately, had no effect on fluid transport by colon sacs, suggesting that the stimulation of fluid transport after the application of angiotensin is not mediated through cyclic AMP. Cycloheximide and puromycin (used at concentrations which block colon protein synthesis by 50–90%) had no effect on fluid transport by control colon sacs, but completely blocked the stimulatory response of the colon to angiotensin. In contrast, actinomycin D (at a concentration which significantly inhibits RNA synthesis) did not affect fluid transport in control or angiotensin-stimulated colon sacs. The results are discussed in relation to the possibility that protein synthesis, at the stage of translation, is involved in the action of angiotensin on fluid transport by the colon.

scholarly journals Optogenetic control of mRNA localization and translation in live cells

2020 ◽  
Author(s):  
Sangkyu Lee ◽  
Won Heo ◽  
Na Kim
Keyword(s):  
Protein Synthesis ◽  
Blue Light ◽  
Protein Interactions ◽  
Mrna Localization ◽  
Spatiotemporal Dynamics ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Protein Clusters ◽  
Specific Mrnas ◽  
Specific Mrna

Abstract Numerous efforts have been made toward the goal of visualizing the spatiotemporal dynamics of single mRNA molecules, yet our capacity for precisely controlling their functions lags behind. Here, we present an optogenetic approach for manipulating the localization and translation of specific mRNAs in live cells. Our technique combines blue light-responsive protein-protein interactions with mRNA visualization modules to robustly and reversibly generate protein clusters that can trap specific mRNA molecules. This sequestration reduces the binding chance of mRNAs with ribosomes, thereby dramatically attenuating protein synthesis

scholarly journals The mechanism of formation of inhibitor-induced ribosome helices in Entamoeba invadens.

1975 ◽  
Vol 65 (3) ◽  
pp. 529-539 ◽  
Author(s):  
T Kusamrarn ◽  
P Sobhon ◽  
G B Bailey
Keyword(s):  
Protein Synthesis ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Aggregate Formation ◽  
Mechanism Of Formation ◽  
Helix Formation ◽  
Aggregation Inhibition ◽  
Chromatoid Bodies ◽  
Entamoeba Invadens ◽  
Slow Growing

Helices andaggregates of helices (chromatoid bodies) composed of ribosomelike particles appear in cysts and slow-growing trophozoites of Entamoeba invadens. We found that similar helix aggregates were formed abundantly in actively growing E. invadens trophozoites treated with a variety of direct or indirect inhibitors of protein synthesis. The inhibitor-induced helices appeared cytochemically and ultrastructurally identical to those seen in cysts. Numerous single helices and small arrays occurred randomly distributed throughout the trophozoite cytoplasm within 15 min after treatment with NaF, which rapidly and completely stopped all nucleic acid and protein synthesis. Cycloheximide (CH), which inhibited protein synthesis as effectively a NaF, stimulated aggregate formation more slowly, and only after a delay of 30-60 min. CH temporarily blocked NaF-stimulated aggregated formation. Aggregation was slowest with actinomycin-D, which strongly inhibited RNA synthesis but depressed protein synthesis only slowly. These results suggested that release of ribosomes from mRNA was required for aggregation. Inhibition by CH was reversible, and aggregates disappeared from CH-treated amebas shortly after they were transferred to inhibitor-free frowth medium. There was no evidence that helices assembled about a structural organizer within the cell or that the process involved metabloc activity. It was concluded that the inhibitor-induced helices were composed of mature, normally functional ribosomes and that helix formation was a spontaneous and reversible consequence of the accumulation withing the cell of free monosomes (or subunits) which were prevented from binding to mRNA.

Embryonale Induktion und Hemmung der Ribonucleinsäure-Synthese durch Actinomycin D

1967 ◽  
Vol 22 (6) ◽  
pp. 649-659 ◽  
Author(s):  
Hildegard Tiedemann ◽  
Jochen Born ◽  
Heinz Tiedemann
Keyword(s):  
Spinal Cord ◽  
Protein Synthesis ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Neural Tissue ◽  
Neural System ◽  
Differential Sensitivity ◽  
Notochordal Cells

Blastoporal lip (presumptive notochord and myotomes) together with adjacent ectoderm (which is induced to form the neural system by the blastoporal lip) from early gastrulae was explanted and treated with 0,5—2,5 µg/ml actinomycin D for periods ranging from two hours to several days. The RNA synthesis is completely inhibited at 2,5 μg/ml Actinomycin D and partially inhibited at 0,5 μg/ml actinomycin D. At 2,5 µg/ml some notochordal cells are still formed, but the differentiation of neural tissue is completely inhibited. At 0,5 μg/ml actinomycin D notochord and muscle are more resistent than the neural tissue.The inducing capacity of the blastoporal lip is not diminished after treatment with actinomycin D. Actinomycin D must therefore impair the competence of the ectoderm to be induced by inhibiting the RNA synthesis.If tissues from tailbuds, which are already differentiating into muscle, notochord and spinal cord are explanted and treated with actinomycin D the differentiation is partially inhibited, but a differential sensitivity of these tissues is not observed.The overall rate of protein synthesis is somewhat enhanced after treatment with 0,5 μg/ml actinomycin D for 2 — 5 hours.

scholarly journals ON THE DIFFERENTIAL CYTOTOXICITY OF ACTINOMYCIN D

1971 ◽  
Vol 50 (3) ◽  
pp. 746-761 ◽  
Author(s):  
Stanley G. Sawicki ◽  
Gabriel C. Godman
Keyword(s):  
Protein Synthesis ◽  
Acute Phase ◽  
Hela Cells ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Differential Susceptibility ◽  
Cell Types ◽  
Cell Degeneration ◽  
L Cells ◽  
Inhibition Of Protein Synthesis

Actinomycin D (AMD) at concentrations that inhibit cellular RNA synthesis by 85% or more causes an acute phase of lethal cell degeneration in HeLa cultures beginning as early as 3 hr after drug exposure, resulting in the nearly complete loss of viable cells by 12 hr. The loss of cells during this acute phase of lethality is closely dose dependent. Vero, WI38, or L cells are not susceptible to this early acute cyto-intoxication by AMD, and may begin to die only after 1–2 days. Differential susceptibility to acute cyto-intoxication by AMD, or other inhibitors of RNA synthesis (daunomycin or nogalamycin), among different types of cultured cells is analogous to that observed in vivo in certain tissues and tumors, and cannot be accounted for by differences in the effect of AMD on RNA, DNA, or protein syntheses, or by the over-all loss of preformed RNA. Actinomycin D in a dose that inhibits RNA synthesis causes an equivalent loss of the prelabeled RNA in all the cell types studied. Inhibition of protein synthesis with streptovitacin A or of DNA synthesis with hydroxyurea does not cause acute lethal injury in HeLa cells as does inhibition of RNA synthesis. Furthermore, since Vero or L cells divide at about the same rate as HeLa cells, no correlation can be drawn between the rate of cell proliferation and susceptibility to the cytotoxicity of AMD. Susceptibile cells are most vulnerable to intoxication by AMD in the G1-S interphase or early S phase. Inhibition of protein synthesis (which protects cells against damage by other agents affecting DNA) does not protect against AMD-induced injury. Although HeLa cells bind more AMD at a given dose than Vero or L cells, the latter cell types, given higher doses, can be made to bind proportionally more AMD without succumbing to acute cyto-intoxication. It is suggested that the differential susceptibility of these cell types to acute poisoning by AMD may reflect differences among various cells in the function or stability of certain RNA species not directly involved in translation whose presence is vital to cells. In HeLa cells, these critical species of RNA are presumed to have a short half-life.

scholarly journals A comparative study of the effect of aflatoxin B1 and actinomycin D on HeLa cells

1969 ◽  
Vol 114 (2) ◽  
pp. 289-298 ◽  
Author(s):  
E. H. Harley ◽  
K. R. Rees ◽  
A. Cohen
Keyword(s):  
Protein Synthesis ◽  
Ribosomal Rna ◽  
Mechanism Of Action ◽  
Aflatoxin B1 ◽  
Hela Cells ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Inhibitory Effect ◽  
Cytotoxic Effects ◽  
Rna Precursor

1. The cytotoxic effects of aflatoxin B1 on HeLa cells were examined and effects of short exposures of the cells to the toxin were found to be reversible. 2. Aflatoxin B1 inhibited the synthesis of both ribosomal and heterodisperse RNA. It is proposed that the toxin's mechanism of action on ribosomal RNA synthesis is related to its inhibitory effect on the maturation of the 45s-ribosomal-RNA precursor. 3. Protein synthesis is inhibited to a greater extent by aflatoxin B1 than by actinomycin D. In contrast with actinomycin D, aflatoxin B1 was shown to disaggregate polyribosomes directly.

Effects of actinomycin D on the lens regenerating system

Development ◽  
1964 ◽  
Vol 12 (4) ◽  
pp. 713-725
Author(s):  
Tuneo Yamada ◽  
Marion E. Roesel
Keyword(s):  
Protein Synthesis ◽  
Early Phase ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Sequence Of Events ◽  
Lens Regeneration

Autoradiographic studies of transformation of iris into lens during Wolffian lens regeneration of adult Triturus viridescens indicate an enhancement of RNA synthesis in the nucleus of the iris cells in the early phase of regeneration (Yamada & Karasaki, 1963; unpublished). Activation of protein synthesis and production of lens antigens follow the enhancement of RNA synthesis (Yamada & Takata, 1963; Takata et al., 1964). This sequence of events suggests the possibility that lens removal elicits synthesis of specific RNA's which in their turn induce synthesis of proteins essential for transformation of the tissue. If this is the case, it should be possible to suppress lens regeneration by subjecting the system to an inhibitor of RNA synthesis. In the present present study the possibility was tested, using actinomycin D as the inhibitor.

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