Stability of histone mRNAs is related to their location in polysomes

1986 ◽  
Vol 64 (2) ◽  
pp. 99-105 ◽  
Author(s):  
R. Curtis Bird ◽  
Frederik A. Jacobs ◽  
Bruce H. Sells
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Histone H4 ◽  
Histone Mrna ◽  
Specific Mechanism ◽  
Histone Mrnas ◽  
Inhibition Of Protein Synthesis ◽  
Rate Of Decay ◽  
Inhibition Of Dna Synthesis ◽  
New Protein

Synthesis of histone mRNAs is closely coupled to DNA synthesis. Following inhibition of DNA synthesis in L6 myoblasts with cytosine arabinoside, a coordinate and exaggerated rate of degradation of histone mRNAs occurs while other mRNAs, encoding ribosomal protein L32 and actin, are unaffected. Inhibition of protein synthesis by puromycin, emetine, or cycloheximide stabilizes histone mRNAs and results in their accumulation. When inhibition of DNA synthesis was followed immediately by inhibition of protein synthesis, the exaggerated rate of decay of the existing subspecies of histone H4 mRNAs was prevented and histone mRNA accumulated. If inhibition of protein synthesis was delayed longer than 3 minutes following inhibition of DNA synthesis, the ability to accumulate H4 mRNAs was lost. Furthermore, new protein synthesis was required to activate the mechanism which specifically destabilized histone mRNA. Puromycin was able to prevent the exaggerated rate of degradation of the various subspecies of H4 mRNA when added up to 15 min after inhibition of DNA synthesis, whereas emetine was effective only when added up to 5 min following inhibition of DNA synthesis. These data suggest that histone H4 mRNAs in polysomes are better targets than those released from polysomes for the specific mechanism which destabilizes histone mRNAs upon inhibition of DNA synthesis.

scholarly journals Investigations of the Mode of Action and Resistance Development of Cadazolid, a New Antibiotic for Treatment of Clostridium difficile Infections

2013 ◽  
Vol 58 (2) ◽  
pp. 901-908 ◽  
Author(s):  
Hans H. Locher ◽  
Patrick Caspers ◽  
Thierry Bruyère ◽  
Susanne Schroeder ◽  
Philippe Pfaff ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Clostridium Difficile ◽  
Dna Synthesis ◽  
Mode Of Action ◽  
Cross Resistance ◽  
Resistance Development ◽  
Content Type ◽  
Inhibition Of Protein Synthesis ◽  
Inhibitor Of Protein Synthesis ◽  
Inhibition Of Dna Synthesis

ABSTRACTCadazolid is a new oxazolidinone-type antibiotic currently in clinical development for the treatment ofClostridium difficile-associated diarrhea. Here, we report investigations on the mode of action and the propensity for spontaneous resistance development inC. difficilestrains. Macromolecular labeling experiments indicated that cadazolid acts as a potent inhibitor of protein synthesis, while inhibition of DNA synthesis was also observed, albeit only at substantially higher concentrations of the drug. Strong inhibition of protein synthesis was also obtained in strains resistant to linezolid, in agreement with low MICs against such strains. Inhibition of protein synthesis was confirmed in coupled transcription/translation assays using extracts from differentC. difficilestrains, including strains resistant to linezolid, while inhibitory effects in DNA topoisomerase assays were weak or not detectable under the assay conditions. Spontaneous resistance frequencies of cadazolid were low in all strains tested (generally <10−10at 2× to 4× the MIC), and in multiple-passage experiments (up to 13 passages) MICs did not significantly increase. Furthermore, no cross-resistance was observed, as cadazolid retained potent activity against strains resistant or nonsusceptible to linezolid, fluoroquinolones, and the new antibiotic fidaxomicin. In conclusion, the data presented here indicate that cadazolid acts primarily by inhibition of protein synthesis, with weak inhibition of DNA synthesis as a potential second mode of action, and suggest a low potential for spontaneous resistance development.

scholarly journals Rapid reversible changes in the rate of histone gene transcription and histone mRNA levels in mouse myeloma cells.

1984 ◽  
Vol 4 (2) ◽  
pp. 351-357 ◽  
Author(s):  
R A Graves ◽  
W F Marzluff
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Gene Transcription ◽  
Histone Gene ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Myeloma Cells ◽  
Histone Mrnas ◽  
Histone Gene Transcription ◽  
Mouse Myeloma

The levels of histone mRNAs are reduced 90 to 95% after treatment of mouse myeloma cells with inhibitors of DNA synthesis which disrupt deoxynucleotide metabolism. In contrast, novobiocin, which inhibits DNA synthesis but does not alter deoxynucleotide metabolism, did not alter histone mRNA levels. Upon reversing the inhibition by fluorodeoxyuridine by feeding with thymidine, histone mRNA levels are restored to control levels within 40 to 60 min. The rate of histone gene transcription is reduced 75 to 80% within 10 min after treatment with fluorodeoxyuridine and increased to control levels within 10 min after refeeding with thymidine. Inhibition of protein synthesis with cycloheximide or puromycin in cells which had been treated with fluorodeoxyuridine resulted in an increase of histone mRNA levels. This was partly due to an increase in the rate of transcription. The data indicate that both transcription and mRNA degradation are linked to deoxynucleotide metabolism. Continued protein synthesis is necessary for maintaining the inhibition of histone gene transcription.

Rapid reversible changes in the rate of histone gene transcription and histone mRNA levels in mouse myeloma cells

1984 ◽  
Vol 4 (2) ◽  
pp. 351-357
Author(s):  
R A Graves ◽  
W F Marzluff
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Gene Transcription ◽  
Histone Gene ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Myeloma Cells ◽  
Histone Mrnas ◽  
Histone Gene Transcription ◽  
Mouse Myeloma

The levels of histone mRNAs are reduced 90 to 95% after treatment of mouse myeloma cells with inhibitors of DNA synthesis which disrupt deoxynucleotide metabolism. In contrast, novobiocin, which inhibits DNA synthesis but does not alter deoxynucleotide metabolism, did not alter histone mRNA levels. Upon reversing the inhibition by fluorodeoxyuridine by feeding with thymidine, histone mRNA levels are restored to control levels within 40 to 60 min. The rate of histone gene transcription is reduced 75 to 80% within 10 min after treatment with fluorodeoxyuridine and increased to control levels within 10 min after refeeding with thymidine. Inhibition of protein synthesis with cycloheximide or puromycin in cells which had been treated with fluorodeoxyuridine resulted in an increase of histone mRNA levels. This was partly due to an increase in the rate of transcription. The data indicate that both transcription and mRNA degradation are linked to deoxynucleotide metabolism. Continued protein synthesis is necessary for maintaining the inhibition of histone gene transcription.

Interferon inhibition of DNA synthesis and cell division

1972 ◽  
Vol 18 (2) ◽  
pp. 145-151 ◽  
Author(s):  
M. V. O'Shaughnessy ◽  
S. H. S. Lee ◽  
K. R. Rozee
Keyword(s):  
Cell Division ◽  
Antiviral Activity ◽  
Dna Synthesis ◽  
S Phase ◽  
Cell Suspensions ◽  
Depression Effect ◽  
Growth Depression ◽  
Synchronized Cells ◽  
Inhibition Of Dna Synthesis ◽  
New Protein

Using monodispersed cell suspensions, interferon preparations were shown to have both a lethal and a growth-depression effect in the same concentration range as that required for antiviral activity. In addition, synchronized cells treated with interferon respond by delaying their normal uptake of thymidine during S phase until after a period during which new protein is synthesized. Puromycin added during this period prevents both the synthesis of this protein and the subsequent synthesis of DNA.

A Comparison of the Effects of Endotoxin Upon Fibroblast Proliferation and Macromolecular Syntheses

1979 ◽  
Vol 58 (6) ◽  
pp. 1634-1639 ◽  
Author(s):  
R.E. Singer ◽  
W.G. Dutton
Keyword(s):  
Escherichia Coli ◽  
Cell Proliferation ◽  
Protein Synthesis ◽  
Dna Synthesis ◽  
L929 Cell ◽  
Cell Protein ◽  
Fibroblast Proliferation ◽  
Proline Incorporation ◽  
Inhibition Of Dna Synthesis

The effects of Escherichia coli endotoxin upon mouse L929 cell proliferation, DNA synthesis, protein synthesis, and proline incorporation were determined. It was found that a level of endotoxin which inhibited cell proliferation prompted a similar inhibition of DNA synthesis and overall cell protein synthesis. In contrast, endotoxin was shown to inhibit incorporation of proline into cell protein to a significantly greater extent.

scholarly journals Long-lived crowded-litter mice have an age-dependent increase in protein synthesis to DNA synthesis ratio and mTORC1 substrate phosphorylation

2014 ◽  
Vol 307 (9) ◽  
pp. E813-E821 ◽  
Author(s):  
Joshua C. Drake ◽  
Danielle R. Bruns ◽  
Frederick F. Peelor ◽  
Laurie M. Biela ◽  
Richard A. Miller ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Litter Size ◽  
Cellular Structures ◽  
Mtorc1 Signaling ◽  
Age Dependent ◽  
Model Protein ◽  
Substrate Phosphorylation ◽  
Mtor Complex 1 ◽  
New Protein

Increasing mouse litter size [crowded litter (CL)] presumably imposes a transient nutrient stress during suckling and extends lifespan through unknown mechanisms. Chronic calorically restricted and rapamycin-treated mice have decreased DNA synthesis and mTOR complex 1 (mTORC1) signaling but maintained protein synthesis, suggesting maintenance of existing cellular structures. We hypothesized that CL would exhibit similar synthetic and signaling responses to other long-lived models and, by comparing synthesis of new protein to new DNA, that insight may be gained into the potential preservation of existing cellular structures in the CL model. Protein and DNA synthesis was assessed in gastroc complex, heart, and liver of 4- and 7-mo CL mice. We also examined mTORC1 signaling in 3- and 7-mo aged animals. Compared with controls, 4-mo CL had greater DNA synthesis in gastroc complex with no differences in protein synthesis or mTORC1 substrate phosphorylation across tissues. Seven-month CL had less DNA synthesis than controls in heart and greater protein synthesis and mTORC1 substrate phosphorylation across tissues. The increased new protein-to-new DNA synthesis ratio suggests that new proteins are synthesized more so in existing cells at 7 mo, differing from 4 mo, in CL vs. controls. We propose that, in CL, protein synthesis shifts from being directed toward new cells (4 mo) to maintenance of existing cellular structures (7 mo), independently of decreased mTORC1.

scholarly journals Regulation of human histone gene expression: kinetics of accumulation and changes in the rate of synthesis and in the half-lives of individual histone mRNAs during the HeLa cell cycle.

1983 ◽  
Vol 3 (4) ◽  
pp. 539-550 ◽  
Author(s):  
N Heintz ◽  
H L Sive ◽  
R G Roeder
Keyword(s):  
Cell Cycle ◽  
Hela Cell ◽  
Dna Synthesis ◽  
Fold Increase ◽  
S Phase ◽  
Histone Mrna ◽  
Mrna Accumulation ◽  
Histone Mrnas ◽  
The Individual ◽  
Kinetics Of

We have analyzed the kinetics of accumulation of each of the individual core histone mRNAs throughout the HeLa cell cycle in cells synchronized by sequential thymidine and aphidicolin treatments. These analyses showed that during the S phase there was a 15-fold increase in the levels of histone mRNAs and that this resulted from both an increased rate of synthesis and a lengthening of the half-life of histone mRNAs. A comparison of the kinetics of accumulation of histone mRNA in the total cellular and nuclear RNA populations suggested an increased transcription rate through the S phase. Within 30 min after the inhibition of DNA synthesis in mid-S phase, the steady-state concentration and the rate of synthesis of histone mRNA each declined to their non-S-phase levels. Reactivation of histone mRNA accumulation could occur even after an extended mid-S-phase block in DNA synthesis. These results suggest that the mechanisms responsible for histone mRNA synthesis are not restricted to the G1/S boundary of the HeLa cell cycle, but can operate whenever DNA synthesis is occurring.

Histone H2A.X gene transcription is regulated differently than transcription of other replication-linked histone genes

1993 ◽  
Vol 13 (2) ◽  
pp. 984-992
Author(s):  
W M Bonner ◽  
C Mannironi ◽  
A Orr ◽  
D R Pilch ◽  
C L Hatch
Keyword(s):  
Protein Synthesis ◽  
Gene Transcription ◽  
Structure Characteristic ◽  
Loop Structure ◽  
Histone Genes ◽  
Histone H2a ◽  
Stem Loop ◽  
Stem Loop Structure ◽  
Inhibition Of Protein Synthesis ◽  
Inhibition Of Dna Synthesis

Histone H2A.X is a replication-independent histone H2A isoprotein species that is encoded by a transcript alternatively processed at the 3' end to yield two mRNAs: a 0.6-kb mRNA ending with the stem-loop structure characteristic of the mRNAs for replication-linked histone species, and a second, polyadenylated 1.6-kb mRNA ending about 1 kb further downstream (C. Mannironi, W. M. Bonner, and C. L. Hatch, Nucleic Acids Res. 17:9113-9126, 1989). Of the two, the 0.6-kb H2A.X stem-loop mRNA predominates in many cell lines, indicating that the presence of two types of mRNA may not completely account for the replication independence of H2A.X protein synthesis. The ambiguity is resolved by the finding that the level of the 0.6-kb H2A.X mRNA is only weakly downregulated during the inhibition of DNA replication and only weakly upregulated during the inhibition of protein synthesis, while the levels of other replication-linked mRNAs are strongly down- or upregulated under these two conditions. Analysis of the nuclear transcription rates of specific H2A genes showed that while the rates of transcription of replication-linked H2A genes decreased substantially during the inhibition of DNA synthesis and increased substantially during the inhibition of protein synthesis, the rate of H2A.X gene transcription decreased slightly under both conditions. These differences in transcriptional regulation between the H2A.X gene and other replication-linked histone genes are sufficient to account for the differences in regulation of their respective stem-loop mRNAs.

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