scholarly journals Use of a cell cycle mutant to delineate the critical period for the control of histone mRNA levels in the mammalian cell cycle.

1984 ◽  
Vol 4 (11) ◽  
pp. 2364-2369 ◽  
Author(s):  
A Artishevsky ◽  
A M Delegeane ◽  
A S Lee
Keyword(s):  
Cell Cycle ◽  
Critical Period ◽  
Fibroblast Cell ◽  
Temporal Analysis ◽  
S Phase ◽  
Temperature Sensitive ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Mrna Accumulation ◽  
Mammalian Cell Cycle

Temporal analysis of DNA replication and histone mRNA accumulation in a hamster fibroblast cell cycle mutant (K12) showed that histone mRNA accumulates periodically during the cell cycle and reaches its highest level in the S phase. The direct correlation between the initiation of DNA synthesis and the accumulation of histone mRNA to high levels in S phase demonstrated the strict interdependence of these two events. Moreover, a critical period necessary for histone mRNA accumulation occurred late in G1 phase. If cells were incubated at the nonpermissive temperature during this critical period, the amount of histone mRNA remained at the basal level. Transcription rate measurements indicated that the triggering of histone mRNA synthesis occurred in late G1 and this mRNA was synthesized at its maximal rate 3 to 5 h before its peak of accumulation. However, if cells were prohibited from synthesizing DNA as a consequence of the temperature-sensitive block in G1, the synthesis of histone mRNA was not initiated.

Use of a cell cycle mutant to delineate the critical period for the control of histone mRNA levels in the mammalian cell cycle

1984 ◽  
Vol 4 (11) ◽  
pp. 2364-2369
Author(s):  
A Artishevsky ◽  
A M Delegeane ◽  
A S Lee
Keyword(s):  
Cell Cycle ◽  
Critical Period ◽  
Fibroblast Cell ◽  
Temporal Analysis ◽  
S Phase ◽  
Temperature Sensitive ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Mrna Accumulation ◽  
Mammalian Cell Cycle

Temporal analysis of DNA replication and histone mRNA accumulation in a hamster fibroblast cell cycle mutant (K12) showed that histone mRNA accumulates periodically during the cell cycle and reaches its highest level in the S phase. The direct correlation between the initiation of DNA synthesis and the accumulation of histone mRNA to high levels in S phase demonstrated the strict interdependence of these two events. Moreover, a critical period necessary for histone mRNA accumulation occurred late in G1 phase. If cells were incubated at the nonpermissive temperature during this critical period, the amount of histone mRNA remained at the basal level. Transcription rate measurements indicated that the triggering of histone mRNA synthesis occurred in late G1 and this mRNA was synthesized at its maximal rate 3 to 5 h before its peak of accumulation. However, if cells were prohibited from synthesizing DNA as a consequence of the temperature-sensitive block in G1, the synthesis of histone mRNA was not initiated.

scholarly journals Regulation of histone mRNA in the unperturbed cell cycle: evidence suggesting control at two posttranscriptional steps.

1991 ◽  
Vol 11 (5) ◽  
pp. 2416-2424 ◽  
Author(s):  
M E Harris ◽  
R Böhni ◽  
M H Schneiderman ◽  
L Ramamurthy ◽  
D Schümperli ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Half Life ◽  
S Phase ◽  
Protein Synthesis Inhibitor ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Synthesis Inhibitor ◽  
Processing Efficiency ◽  
Histone 3 ◽  
Cycle Regulation

The levels of histone mRNA increase 35-fold as selectively detached mitotic CHO cells progress from mitosis through G1 and into S phase. Using an exogenous gene with a histone 3' end which is not sensitive to transcriptional or half-life regulation, we show that 3' processing is regulated as cells progress from G1 to S phase. The half-life of histone mRNA is similar in G1- and S-phase cells, as measured after inhibition of transcription by actinomycin D (dactinomycin) or indirectly after stabilization by the protein synthesis inhibitor cycloheximide. Taken together, these results suggest that the change in histone mRNA levels between G1- and S-phase cells must be due to an increase in the rate of biosynthesis, a combination of changes in transcription rate and processing efficiency. In G2 phase, there is a rapid 35-fold decrease in the histone mRNA concentration which our results suggest is due primarily to an altered stability of histone mRNA. These results are consistent with a model for cell cycle regulation of histone mRNA levels in which the effects on both RNA 3' processing and transcription, rather than alterations in mRNA stability, are the major mechanisms by which low histone mRNA levels are maintained during G1.

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.

scholarly journals Phosphorylation of Threonine 61 by Cyclin A/Cdk1 Triggers Degradation of Stem-Loop Binding Protein at the End of S Phase

2008 ◽  
Vol 28 (14) ◽  
pp. 4469-4479 ◽  
Author(s):  
M. Murat Koseoglu ◽  
Lee M. Graves ◽  
William F. Marzluff
Keyword(s):  
Cell Cycle ◽  
Fusion Proteins ◽  
Regulatory Mechanism ◽  
Binding Protein ◽  
Cyclin A ◽  
S Phase ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Stem Loop

ABSTRACT Histone mRNA levels are cell cycle regulated, and a major regulatory mechanism is restriction of stem-loop binding protein (SLBP) to S phase. Degradation of SLBP at the end of S phase results in cessation of histone mRNA biosynthesis, preventing accumulation of histone mRNA until SLBP is synthesized just before entry into the next S phase. Degradation of SLBP requires an SFTTP (58 to 62) and KRKL (95 to 98) sequence, which is a putative cyclin binding site. A fusion protein with the 58-amino-acid sequence of SLBP (amino acids 51 to 108) fused to glutathione S-transferase (GST) is sufficient to mimic SLBP degradation at late S phase. Using GST-SLBP fusion proteins as a substrate, we show that cyclin A/Cdk1 phosphorylates Thr61. Furthermore, knockdown of Cdk1 by RNA interference stabilizes SLBP at the end of S phase. Phosphorylation of Thr61 is necessary for subsequent phosphorylation of Thr60 by CK2 in vitro. Inhibitors of CK2 also prevent degradation of SLBP at the end of S phase. Thus, phosphorylation of Thr61 by cyclin A/Cdk1 primes phosphorylation of Thr60 by CK2 and is responsible for initiating SLBP degradation. We conclude that the increase in cyclin A/Cdk1 activity at the end of S phase triggers degradation of SLBP at S/G2.

Regulation of histone mRNA in the unperturbed cell cycle: evidence suggesting control at two posttranscriptional steps

1991 ◽  
Vol 11 (5) ◽  
pp. 2416-2424
Author(s):  
M E Harris ◽  
R Böhni ◽  
M H Schneiderman ◽  
L Ramamurthy ◽  
D Schümperli ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Half Life ◽  
S Phase ◽  
Protein Synthesis Inhibitor ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Synthesis Inhibitor ◽  
Processing Efficiency ◽  
Histone 3 ◽  
Cycle Regulation

The levels of histone mRNA increase 35-fold as selectively detached mitotic CHO cells progress from mitosis through G1 and into S phase. Using an exogenous gene with a histone 3' end which is not sensitive to transcriptional or half-life regulation, we show that 3' processing is regulated as cells progress from G1 to S phase. The half-life of histone mRNA is similar in G1- and S-phase cells, as measured after inhibition of transcription by actinomycin D (dactinomycin) or indirectly after stabilization by the protein synthesis inhibitor cycloheximide. Taken together, these results suggest that the change in histone mRNA levels between G1- and S-phase cells must be due to an increase in the rate of biosynthesis, a combination of changes in transcription rate and processing efficiency. In G2 phase, there is a rapid 35-fold decrease in the histone mRNA concentration which our results suggest is due primarily to an altered stability of histone mRNA. These results are consistent with a model for cell cycle regulation of histone mRNA levels in which the effects on both RNA 3' processing and transcription, rather than alterations in mRNA stability, are the major mechanisms by which low histone mRNA levels are maintained during G1.

Complementation of G1-phase variants of a mammalian cell cycle

1979 ◽  
Vol 35 (1) ◽  
pp. 53-58
Author(s):  
P.M. Naha
Keyword(s):  
Cell Cycle ◽  
Cell Fusion ◽  
Mammalian Cell ◽  
S Phase ◽  
G1 Phase ◽  
Temperature Sensitive ◽  
Multinucleated Cells ◽  
Mammalian Cell Cycle ◽  
Complementation Groups ◽  
Phase Variants

Complementation between temperature-sensitive (ts) variants of Balb/c-3T3 defective in the G1 phase of its cell cycle was measured in the [3H]thymidine-labeling indices of the multinucleated cells during incubation at the restricted temperature (38 degrees C) following cell fusion. One ts variant from each group along the length of the G1 phase was tested for complementation. Varying degrees of complementation were observed between the 4 ts variants tested, judging by the time of entry into S-phase and the degree of synchrony attained. At least 3 complementation groups were discernible.

scholarly journals Stem-Loop Binding Protein, the Protein That Binds the 3′ End of Histone mRNA, Is Cell Cycle Regulated by Both Translational and Posttranslational Mechanisms

2000 ◽  
Vol 20 (12) ◽  
pp. 4188-4198 ◽  
Author(s):  
Michael L. Whitfield ◽  
Lian-Xing Zheng ◽  
Amy Baldwin ◽  
Tomohiko Ohta ◽  
Myra M. Hurt ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Regulation ◽  
Binding Protein ◽  
S Phase ◽  
Mrna Processing ◽  
Mrna Levels ◽  
Histone Mrna ◽  
Stem Loop ◽  
Histone Mrnas ◽  
Cycle Regulation

ABSTRACT The expression of the replication-dependent histone mRNAs is tightly regulated during the cell cycle. As cells progress from G1 to S phase, histone mRNA levels increase 35-fold, and they decrease again during G2 phase. Replication-dependent histone mRNAs are the only metazoan mRNAs that lack polyadenylated tails, ending instead in a conserved stem-loop. Much of the cell cycle regulation is posttranscriptional and is mediated by the 3′ stem-loop. A 31-kDa stem-loop binding protein (SLBP) binds the 3′ end of histone mRNA. The SLBP is necessary for pre-mRNA processing and accompanies the histone mRNA to the cytoplasm, where it is a component of the histone messenger RNP. We used synchronous CHO cells selected by mitotic shakeoff and HeLa cells synchronized at the G1/S or the M/G1 boundary to study the regulation of SLBP during the cell cycle. In each system the amount of SLBP is regulated during the cell cycle, increasing 10- to 20-fold in the late G1 and then decreasing in the S/G2border. SLBP mRNA levels are constant during the cell cycle. SLBP is regulated at the level of translation as cells progress from G1 to S phase, and the protein is rapidly degraded as they progress into G2. Regulation of SLBP may account for the posttranscriptional component of the cell cycle regulation of histone mRNA.

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.

In vitro evidence that LHRH stimulates the recruitment of prolactin mRNA-expressing cells during the postnatal period in the rat

1994 ◽  
Vol 12 (1) ◽  
pp. 107-118 ◽  
Author(s):  
A Van Bael ◽  
R Huygen ◽  
B Himpens ◽  
C Denef
Keyword(s):  
Cell Cycle ◽  
Cell Population ◽  
Blot Hybridization ◽  
Postnatal Period ◽  
S Phase ◽  
Female Rats ◽  
Mrna Levels ◽  
Dot Blot ◽  
Total Cell Population

ABSTRACT We have studied the effect of LHRH and neuropeptide Y (NPY) on prolactin (PRL) mRNA levels in pituitary reaggregate cell cultures from 14-day-old female rats, by means of in situ hybridization and Northern blot analysis. As estimated by computer-image analysis, addition of LHRH on day 5 in culture for 40 h resulted in a 37% increase in the total cytoplasmic areas of cells containing PRL mRNA, visualized using a digoxigenin-labelled PRL cRNA. The size of individual PRL-expressing cells was not influenced, nor was the content of PRL mRNA per cell. A similar effect of LHRH was found by dot blot hybridization of extracted RNA. PRL mRNA levels were not affected by NPY. LHRH induced a 29% increase in the number of PRL mRNA-expressing cells processing through the S phase of the cell cycle, visualized by the incorporation of [3H]thymidine ([3H]T) into DNA over 16 h. The fraction of [3H]T-labelled cells was 10–12% of the total cell population. NPY did not influence the number of [3H]T-positive cells expressing PRL mRNA, but completely blocked the effect of LHRH on the latter population. The present data suggest that LHRH, probably via a paracrine action of gonadotrophs, stimulates the recruitment of new lactotrophs, an action which is negatively modulated by NPY. Since the magnitude of this effect was the same in the total pituitary cell population as in cells processing through the S phase of the cell cycle and presumably mitosis, recruitment of lactotrophs seems to be based on differentiation of progenitor or immature cells into PRL-expressing cells, rather than on a mitogenic action on pre-existing lactotrophs alone.

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