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.

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 Stem-Loop Binding Protein (SLBP) on Two Threonines Triggers Degradation of SLBP, the Sole Cell Cycle-Regulated Factor Required for Regulation of Histone mRNA Processing, at the End of S Phase

2003 ◽  
Vol 23 (5) ◽  
pp. 1590-1601 ◽  
Author(s):  
Lianxing Zheng ◽  
Zbigniew Dominski ◽  
Xiao-Cui Yang ◽  
Phillip Elms ◽  
Christy S. Raska ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Amino Acids ◽  
Posttranscriptional Regulation ◽  
Binding Protein ◽  
S Phase ◽  
Mrna Processing ◽  
Histone Mrna ◽  
Stem Loop ◽  
Histone Mrnas ◽  
Nuclear Extracts

ABSTRACT The replication-dependent histone mRNAs, the only eukaryotic mRNAs that do not have poly(A) tails, are present only in S-phase cells. Coordinate posttranscriptional regulation of histone mRNAs is mediated by the stem-loop at the 3′ end of histone mRNAs. The protein that binds the 3′ end of histone mRNA, stem-loop binding protein (SLBP), is required for histone pre-mRNA processing and is involved in multiple aspects of histone mRNA metabolism. SLBP is also regulated during the cell cycle, accumulating as cells enter S phase and being rapidly degraded as cells exit S phase. Mutation of any residues in a TTP sequence (amino acids 60 to 62) or mutation of a consensus cyclin binding site (amino acids 99 to 104) stabilizes SLBP in G2 and mitosis. These two threonines are phosphorylated in late S phase, as determined by mass spectrometry (MS) of purified SLBP from late S-phase cells, triggering SLBP degradation. Cells that express a stable SLBP still degrade histone mRNA at the end of S phase, demonstrating that degradation of SLBP is not required for histone mRNA degradation. Nuclear extracts from G1 and G2 cells are deficient in histone pre-mRNA processing, which is restored by addition of recombinant SLBP, indicating that SLBP is the only cell cycle-regulated factor required for histone pre-mRNA processing.

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.

Cell cycle regulation of mouse H3 histone mRNA metabolism

1984 ◽  
Vol 4 (1) ◽  
pp. 123-132
Author(s):  
R B Alterman ◽  
S Ganguly ◽  
D H Schulze ◽  
W F Marzluff ◽  
C L Schildkraut ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Steady State ◽  
Dna Synthesis ◽  
In Vitro Transcription ◽  
S Phase ◽  
Gene Probe ◽  
Histone Mrna ◽  
Mrna Content

The mechanisms responsible for the periodic accumulation and decay of histone mRNA in the mammalian cell cycle were investigated in mouse erythroleukemia cells, using a cloned mouse H3 histone gene probe that hybridizes with most or all H3 transcripts. Exponentially growing cells were fractionated into cell cycle-specific stages by centrifugal elutriation, a method for purifying cells at each stage of the cycle without the use of treatments that arrest growth. Measurements of H3 histone mRNA content throughout the cell cycle show that the mRNA accumulates gradually during S phase, achieving its highest value in mid-S phase when DNA synthesis is maximal. The mRNA content then decreases as cells approach G2. These results demonstrate that the periodic synthesis of histones during S phase is due to changes in the steady-state level of histone mRNA. They are consistent with the conventional view in which histone synthesis is regulated coordinately with DNA synthesis in the cell cycle. The periodic accumulation and decay of H3 histone mRNA appear to be controlled primarily by changes in the rate of appearance of newly synthesized mRNA in the cytoplasm, determined by pulse-labeling whole cells with [3H]uridine. Measurements of H3 mRNA turnover by pulse-chase experiments with cells in S and G2 did not provide evidence for changes in the cytoplasmic stability of the mRNA during the period of its decay in late S and G2. Furthermore, transcription measurements carried out by brief pulse-labeling in vivo and by in vitro transcription in isolated nuclei indicate that the rate of H3 gene transcription changes to a much smaller extent than the steady-state levels of the mRNA or the appearance of newly synthesized mRNA in the cytoplasm. The results suggest that post-transcriptional processes make an important contribution to the periodic accumulation and decay of histone mRNA and that these processes may operate within the nucleus.

scholarly journals Distinct replication-independent and -dependent phases of histone gene expression during the Physarum cell cycle.

1987 ◽  
Vol 7 (5) ◽  
pp. 1933-1937 ◽  
Author(s):  
J J Carrino ◽  
V Kueng ◽  
R Braun ◽  
T G Laffler
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Dna Replication ◽  
Dna Synthesis ◽  
S Phase ◽  
Histone Gene ◽  
Histone Mrna ◽  
Histone Gene Expression ◽  
Tightly Coupled ◽  
G2 Phase

During the S phase of the cell cycle, histone gene expression and DNA replication are tightly coupled. In mitotically synchronous plasmodia of the myxomycete Physarum polycephalum, which has no G1 phase, histone mRNA synthesis begins in mid-G2 phase. Although histone gene transcription is activated in the absence of significant DNA synthesis, our data demonstrate that histone gene expression became tightly coupled to DNA replication once the S phase began. There was a transition from the replication-independent phase to the replication-dependent phase of histone gene expression. During the first phase, histone mRNA synthesis appears to be under direct cell cycle control; it was not coupled to DNA replication. This allowed a pool of histone mRNA to accumulate in late G2 phase, in anticipation of future demand. The second phase began at the end of mitosis, when the S phase began, and expression became homeostatically coupled to DNA replication. This homeostatic control required continuing protein synthesis, since cycloheximide uncoupled transcription from DNA synthesis. Nuclear run-on assays suggest that in P. polycephalum this coupling occurs at the level of transcription. While histone gene transcription appears to be directly switched on in mid-G2 phase and off at the end of the S phase by cell cycle regulators, only during the S phase was the level of transcription balanced with the rate of DNA synthesis.

scholarly journals Cell cycle regulation of mouse H3 histone mRNA metabolism.

1984 ◽  
Vol 4 (1) ◽  
pp. 123-132 ◽  
Author(s):  
R B Alterman ◽  
S Ganguly ◽  
D H Schulze ◽  
W F Marzluff ◽  
C L Schildkraut ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Steady State ◽  
Dna Synthesis ◽  
In Vitro Transcription ◽  
S Phase ◽  
Gene Probe ◽  
Histone Mrna ◽  
Mrna Content

The mechanisms responsible for the periodic accumulation and decay of histone mRNA in the mammalian cell cycle were investigated in mouse erythroleukemia cells, using a cloned mouse H3 histone gene probe that hybridizes with most or all H3 transcripts. Exponentially growing cells were fractionated into cell cycle-specific stages by centrifugal elutriation, a method for purifying cells at each stage of the cycle without the use of treatments that arrest growth. Measurements of H3 histone mRNA content throughout the cell cycle show that the mRNA accumulates gradually during S phase, achieving its highest value in mid-S phase when DNA synthesis is maximal. The mRNA content then decreases as cells approach G2. These results demonstrate that the periodic synthesis of histones during S phase is due to changes in the steady-state level of histone mRNA. They are consistent with the conventional view in which histone synthesis is regulated coordinately with DNA synthesis in the cell cycle. The periodic accumulation and decay of H3 histone mRNA appear to be controlled primarily by changes in the rate of appearance of newly synthesized mRNA in the cytoplasm, determined by pulse-labeling whole cells with [3H]uridine. Measurements of H3 mRNA turnover by pulse-chase experiments with cells in S and G2 did not provide evidence for changes in the cytoplasmic stability of the mRNA during the period of its decay in late S and G2. Furthermore, transcription measurements carried out by brief pulse-labeling in vivo and by in vitro transcription in isolated nuclei indicate that the rate of H3 gene transcription changes to a much smaller extent than the steady-state levels of the mRNA or the appearance of newly synthesized mRNA in the cytoplasm. The results suggest that post-transcriptional processes make an important contribution to the periodic accumulation and decay of histone mRNA and that these processes may operate within the nucleus.

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.

scholarly journals Changes in the stability of a human H3 histone mRNA during the HeLa cell cycle

1991 ◽  
Vol 11 (1) ◽  
pp. 544-553
Author(s):  
T D Morris ◽  
L A Weber ◽  
E Hickey ◽  
G S Stein ◽  
J L Stein
Keyword(s):  
Cell Cycle ◽  
Hela Cell ◽  
Half Life ◽  
Fusion Gene ◽  
S Phase ◽  
Histone Mrna ◽  
Histone Protein ◽  
Heat Induction ◽  
Hela Cell Lines ◽  
The Stability

A major component of the regulation of histone protein synthesis during the cell cycle is the modulation of the half-life of histone mRNA. We have uncoupled transcriptional and posttranscriptional regulation by using a Drosophila hsp70-human H3 histone fusion gene that produces a marked human H3 histone mRNA upon heat induction. Transcription of this gene can be switched on and off by raising and lowering cell culture temperatures, respectively. HeLa cell lines containing stably integrated copies of the fusion gene were synchronized by double thymidine block. Distinct populations of H3 histone mRNA were produced by heat induction in early S-phase, late S-phase, or G2-phase cells, and the stability of the induced H3 histone mRNA was measured. The H3 histone mRNA induced during early S phase decayed with a half-life of 110 min, whereas the same transcript induced during late S phase had a half-life of 10 to 15 min. The H3 histone mRNA induced in non-S-phase cells is more stable than that induced in late S phase, with a half-life of 40 min. Thus, the stability of histone mRNA is actively regulated throughout the cell cycle. Our results are consistent with an autoregulatory model in which the stability of histone mRNA is determined by the level of free histone protein in the cytoplasm.

Distinct replication-independent and -dependent phases of histone gene expression during the Physarum cell cycle

1987 ◽  
Vol 7 (5) ◽  
pp. 1933-1937
Author(s):  
J J Carrino ◽  
V Kueng ◽  
R Braun ◽  
T G Laffler
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Dna Replication ◽  
Dna Synthesis ◽  
S Phase ◽  
Histone Gene ◽  
Histone Mrna ◽  
Histone Gene Expression ◽  
Tightly Coupled ◽  
G2 Phase

During the S phase of the cell cycle, histone gene expression and DNA replication are tightly coupled. In mitotically synchronous plasmodia of the myxomycete Physarum polycephalum, which has no G1 phase, histone mRNA synthesis begins in mid-G2 phase. Although histone gene transcription is activated in the absence of significant DNA synthesis, our data demonstrate that histone gene expression became tightly coupled to DNA replication once the S phase began. There was a transition from the replication-independent phase to the replication-dependent phase of histone gene expression. During the first phase, histone mRNA synthesis appears to be under direct cell cycle control; it was not coupled to DNA replication. This allowed a pool of histone mRNA to accumulate in late G2 phase, in anticipation of future demand. The second phase began at the end of mitosis, when the S phase began, and expression became homeostatically coupled to DNA replication. This homeostatic control required continuing protein synthesis, since cycloheximide uncoupled transcription from DNA synthesis. Nuclear run-on assays suggest that in P. polycephalum this coupling occurs at the level of transcription. While histone gene transcription appears to be directly switched on in mid-G2 phase and off at the end of the S phase by cell cycle regulators, only during the S phase was the level of transcription balanced with the rate of DNA synthesis.

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.

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