Tubulin gene expression in the Chlamydomonas reinhardtii cell cycle: elimination of environmentally induced artifacts and the measurement of tubulin mRNA levels

1988 ◽  
Vol 89 (3) ◽  
pp. 397-403
Author(s):  
D.S. Nicholl ◽  
J.A. Schloss ◽  
P.C. John
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Division ◽  
Chlamydomonas Reinhardtii ◽  
Environmental Conditions ◽  
Mrna Levels ◽  
Tubulin Gene ◽  
Mrna Accumulation ◽  
Tubulin Mrna ◽  
Flagellar Proteins

To investigate the involvement of tubulin gene expression in controlling cell division events in Chlamydomonas reinhardtii we have measured tubulin mRNA levels during the cell cycle under different environmental conditions. In C. reinhardtii cells grown under the synchronizing conditions of 14 h of light followed by 10 h of darkness, mRNAs for tubulin and associated flagellar proteins were found to accumulate periodically with a peak just prior to cell division. This was not seen when previously synchronized cells were transferred to constant environmental conditions in a turbidostat, suggesting that dramatic changes in tubulin mRNA levels are not required for successful completion of the cell cycle. A hypothesis to explain the patterns of tubulin mRNA accumulation found under different environmental conditions is presented.

Gene expression of human DNA polymerase alpha during cell proliferation and the cell cycle

1988 ◽  
Vol 8 (11) ◽  
pp. 5016-5025
Author(s):  
A F Wahl ◽  
A M Geis ◽  
B H Spain ◽  
S W Wong ◽  
D Korn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Steady State ◽  
Dna Polymerase ◽  
Transformed Cells ◽  
Mrna Levels ◽  
Dna Polymerase Alpha ◽  
Human Dna ◽  
Alpha Gene

We studied the expression of the human DNA polymerase alpha gene during cell proliferation, during cell progression through the cell cycle, and in transformed cells compared with normal cells. During the activation of quiescent cells (G0 phase) to proliferate (G1/S phases), the steady-state mRNA levels, rate of synthesis of nascent polymerase protein, and enzymatic activity in vitro exhibited a substantial and concordant increase prior to the peak of in vivo DNA synthesis. In transformed cells, the respective values were amplified greater than 10-fold. In actively growing cells separated into discrete stages of the cell cycle by counterflow elutriation or by mitotic shakeoff, levels of steady-state transcripts, translation rates, and enzymatic activities of polymerase alpha were constitutively and concordantly expressed at all stages of the cell cycle, with only a moderate elevation prior to the S phase and a slight decline in the G2 phase. These findings support the conclusion that the regulation of human DNA polymerase alpha gene expression is at the transcriptional level and strongly suggest that the regulatory mechanisms that are operative during the entrance of a cell into the mitotic cycle are fundamentally different from those that modulate polymerase alpha expression in continuously cycling cells.

Sequences controlling transcription of the Chlamydomonas reinhardtii beta 2-tubulin gene after deflagellation and during the cell cycle

1994 ◽  
Vol 14 (8) ◽  
pp. 5165-5174
Author(s):  
J P Davies ◽  
A R Grossman
Keyword(s):  
Cell Cycle ◽  
Chlamydomonas Reinhardtii ◽  
Transcription Initiation ◽  
Chimeric Gene ◽  
Transcriptional Level ◽  
Sequence Motif ◽  
Tubulin Gene ◽  
Rich Region ◽  
Beta 2 ◽  
Encoding Genes

In Chlamydomonas reinhardtii, transcripts from the beta 2-tubulin gene (tubB2), as well as those from other tubulin-encoding genes, accumulate immediately after flagellar excision as well as at a specific time in the cell cycle. Control of tubB2 transcript accumulation following deflagellation is regulated, at least partially, at the transcriptional level. We have fused the tubB2 promoter to the arylsulfatase (ars) reporter gene, introduced this construct into C. reinhardtii, and compared expression of the chimeric gene with that of the endogenous tubB2 gene. After flagellar excision, transcripts from the tubB2/ars chimeric gene accumulate with kinetics similar to those of transcripts from the endogenous tubB2 gene. The tubB2/ars transcripts also accumulate in a cell cycle-specific manner; however, chimeric transcripts are more abundant earlier in the cell cycle than the endogenous tubB2 transcripts. To elucidate transcriptional control of tubB2, we have mutated or removed sequences in the tubB2 promoter and examined the effect on transcription. The tubB2 promoter shares features with the promoters of other tubulin-encoding genes; these include a GC-rich region between the TATA box and the transcription initiation site and multiple copies of a 10-bp sequence motif that we call the tub box. The tubB2 gene contains seven tub box motifs. Changing the GC-rich region to an AT-rich region or removing three of the seven tub box motifs did not significantly affect transcription of the chimeric gene. However, removing four or five tub box motifs prevented increased transcription following deflagellation and diminished cell cycle-regulated transcription from the tubB2 promoter.

scholarly journals Alternative Splicing During the Chlamydomonasreinhardtii Cell Cycle

2020 ◽  
Vol 10 (10) ◽  
pp. 3797-3810
Author(s):  
Manishi Pandey ◽  
Gary D. Stormo ◽  
Susan K. Dutcher
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Alternative Splicing ◽  
Chlamydomonas Reinhardtii ◽  
Intron Retention ◽  
Exon Skipping ◽  
Dark Phase ◽  
Periodic Patterns ◽  
Genome Wide ◽  
Splicing Pattern

Genome-wide analysis of transcriptome data in Chlamydomonas reinhardtii shows periodic patterns in gene expression levels when cultures are grown under alternating light and dark cycles so that G1 of the cell cycle occurs in the light phase and S/M/G0 occurs during the dark phase. However, alternative splicing, a process that enables a greater protein diversity from a limited set of genes, remains largely unexplored by previous transcriptome based studies in C. reinhardtii. In this study, we used existing longitudinal RNA-seq data obtained during the light-dark cycle to investigate the changes in the alternative splicing pattern and found that 3277 genes (19.75% of 17,746 genes) undergo alternative splicing. These splicing events include Alternative 5′ (Alt 5′), Alternative 3′ (Alt 3′) and Exon skipping (ES) events that are referred as alternative site selection (ASS) events and Intron retention (IR) events. By clustering analysis, we identified a subset of events (26 ASS events and 10 IR events) that show periodic changes in the splicing pattern during the cell cycle. About two-thirds of these 36 genes either introduce a pre-termination codon (PTC) or introduce insertions or deletions into functional domains of the proteins, which implicate splicing in altering gene function. These findings suggest that alternative splicing is also regulated during the Chlamydomonas cell cycle, although not as extensively as changes in gene expression. The longitudinal changes in the alternative splicing pattern during the cell cycle captured by this study provides an important resource to investigate alternative splicing in genes of interest during the cell cycle in Chlamydomonas reinhardtii and other eukaryotes.

Gene expression after resumption of development of Artemia franciscana cryptobiotic embryos

1994 ◽  
Vol 72 (3-4) ◽  
pp. 78-83 ◽  
Author(s):  
Ricardo Escalante ◽  
Alberto García-Sáez ◽  
Maria-Asunción Ortega ◽  
Leandro Sastre
Keyword(s):  
Gene Expression ◽  
Time Course ◽  
Developmental Stages ◽  
Artemia Franciscana ◽  
Mrna Levels ◽  
Muscle Actin ◽  
Mrna Accumulation ◽  
Α Subunit ◽  
Steady State Levels ◽  
Cyst Development

The steady-state levels of six different mRNAs have been studied during Artemia franciscana development. Some of these mRNAs are present in the cryptobiotic cyst, like those coding for cytoplasmic actins, sarcoplasmic/endoplasmic reticulum Ca2+-ATPase, and the Na+,K+-ATPase α-subunit isoform coded by the clone pArATNa136. The expression of these mRNAs is markedly induced during cyst development. A small increase in mRNA levels can be observed for some genes at very early stages of development (2 h). The main increase is observed between 4 and 16 h of development for all these genes, although the time course of mRNA accumulation is different for each one of the genes studied. Some other genes, like those coding for muscle actin (actin 3) or the Na+,K+-ATPase α-subunit isoform coded by the cDNA clone α2850, are not expressed in the cyst before resumption of development and their expression is induced after 10 or 6 h of development, respectively. These data on the kinetic of mRNA accumulation provide the information required to determine transcriptionally active developmental stages, necessary to study in more detail the mechanisms of transcriptional regulation during activation of cryptobiotic cysts and resumption of embryonic development.Key words: Artemia, gene expression, actin, Na,K-ATPase, Ca2+-ATPase.

Hematopoietic Transcriptional Regulation At The Mitosis-G1 Transition

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2440-2440
Author(s):  
Chris C.S. Hsiung ◽  
Arjun Raj ◽  
Gerd A. Blobel
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Gene Regulation ◽  
Cell Division ◽  
Rna Polymerase Ii ◽  
Single Molecule ◽  
Conflicts Of Interest ◽  
Specific Cell ◽  
Developmental Gene ◽  
Developmental Gene Regulation

Abstract Normal hematopoiesis involves the coordination of cell division and gene expression to produce physiologically appropriate cell numbers of various developmental stages across lineages. While studies have demonstrated intricate links between cell cycle progression and developmental gene regulation -- two cellular programs whose concomitant dysregulation is central to many malignant and non-malignant hematologic diseases -- researchers currently lack clear, general principles of how intrinsic properties of cell division could influence developmental gene regulation. In each round of division, mitosis imposes a striking disruption to gene expression: the nucleus is disassembled, bulk RNA synthesis ceases, and the transcription machinery and most transcription factors -- including repressive complexes -- are evicted from mitotic chromatin. Since hematopoietic lineage fidelity often requires the continued presence of repressive complexes to inhibit expression of developmentally inappropriate genes, we hypothesized that such repression may be inefficient during a narrow window immediately post-mitosis, resulting in transient aberrant transcription in a probabilistic manner. We tested for the presence of transient post-mitotic aberrant transcription at genes whose repression is known to depend on continued occupancy of repressive complexes. We used an experimentally tractable cell line, G1E cells, a rapidly dividing model of lineage-committed murine pro-erythroblasts that genetically lack the erythroid master regulator Gata1. Transduction with a Gata1-estrogen receptor fusion construct and treatment with estradiol restores Gata1 function, leading to recapitulation of early erythroid maturation events, including rapid repression of stemness-associated genes, such as Gata2 and c-Kit. We examined in fine temporal detail the post-mitotic transcriptional behavior of Gata2, c-Kit and other genes using population-based assays facilitated by drug-mediated cell cycle synchronization. In addition, we bypassed the use of synchronization drugs and their associated potential experimental artifacts by developing novel complementary methods to study the relationship between cell cycle status and transcription in asynchronous populations: 1. We harnessed single-molecule RNA fluorescence in situ hybridization technology to quantitatively assess transcription in individual cells at various cell cycle stages, and 2. We adapted a fluorescent protein cell cycle reporter to separate, using fluorescence-activated cell sorting, subpopulations of specific cell cycle stages for epigenomic and transcriptomic analyses. Together, our results revealed a post-mitotic pulse of increased RNA polymerase II recruitment and transcript synthesis most clearly exhibited by Gata2, c-Kit, and other genes whose repression is known to depend on co-repressor complexes in these cells. Our results support the notion that the mitosis-G1 transition presents a window of transcriptional plasticity. We are beginning to explore how this property of post-mitotic transcriptional control applies to hematopoietic cell types across the developmental spectrum and could contribute to functionally important variations in gene expression, such as in stem cell lineage commitment, experimental reprogramming, and non-genetic heterogeneity in malignancy. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Dysregulation of Cyclin E Gene Expression in Human Cytomegalovirus-Infected Cells Requires Viral Early Gene Expression and Is Associated with Changes in the Rb-Related Protein p130

2000 ◽  
Vol 74 (9) ◽  
pp. 4192-4206 ◽  
Author(s):  
Anita K. McElroy ◽  
Roopashree S. Dwarakanath ◽  
Deborah H. Spector
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Human Cytomegalovirus ◽  
Cyclin E ◽  
Early Gene ◽  
Immediate Early ◽  
Mrna Levels ◽  
Early Gene Expression ◽  
Cell Extracts ◽  
Infected Cells

ABSTRACT We have previously shown that many cell cycle regulatory gene products are markedly affected by infection of primary fibroblasts with human cytomegalovirus (HCMV) (F. M. Jault, J. M. Jault, F. Ruchti, E. A. Fortunato, C. Clark, J. Corbeil, D. D. Richman, and D. H. Spector, J. Virol. 69:6697–6704, 1995). One of these proteins, cyclin E, is a key determinant of cell cycle progression during G1, and its mRNA levels are significantly increased in HCMV-infected fibroblasts (B. S. Salvant, E. A. Fortunato, and D. H. Spector, J. Virol. 72:3729–3741, 1998). To determine the molecular basis of this effect, we have examined the events that occur at the endogenous cyclin E promoter during the course of infection. In vivo dimethyl sulfate footprinting of the cyclin E promoter revealed several regions of protection and hypersensitivity that were unique to infected cells. In accord with this observation, we find that the virus-induced cyclin E transcripts initiate downstream of the start site identified in mock-infected cells, in regions where these newly appearing protected and hypersensitive sites occur. Viral gene expression is required for this induction. However, the viral immediate-early proteins IE1-72 and IE2-86, either alone or in combination, cannot induce expression of the endogenous cyclin E. The virus must progress past the immediate-early phase and express an early gene product(s) for activation of cyclin E expression. Moreover, IE1-72 does not appear to be required, as infection of cells with an HCMV mutant containing a deletion in the IE1-72 gene leads to full upregulation of cyclin E expression. Using electrophoretic mobility shift assays with infected cell extracts and a region of the cyclin E promoter that includes two previously defined E2F sites as the probe, we detected the appearance of an infection-specific banding pattern. One of the infection-specific bands contained the proteins E2F-4, DP-1, and p130, which were maintained in the infected cells as uniquely phosphorylated species. These results suggest that an altered E2F-4–DP-1–p130 complex along with viral early gene expression may play a role in the transcriptional regulation of cyclin E mRNA during HCMV infection.

scholarly journals The Gene Expression and Enzyme Activity of Plant 3-Deoxy-D-Manno-2-Octulosonic Acid-8-Phosphate Synthase Are Preferentially Associated with Cell Division in a Cell Cycle-Dependent Manner

2003 ◽  
Vol 133 (1) ◽  
pp. 348-360 ◽  
Author(s):  
Frédéric Delmas ◽  
Johann Petit ◽  
Jérôme Joubès ◽  
Martial Séveno ◽  
Thomas Paccalet ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Enzyme Activity ◽  
Cell Division ◽  
Dependent Manner ◽  
A Cell ◽  
Cycle Dependent ◽  
Phosphate Synthase

scholarly journals Clusterin production in the obstructed rabbit kidney: correlations with loss of renal function.

1992 ◽  
Vol 3 (5) ◽  
pp. 1163-1171
Author(s):  
P N Schlegel ◽  
G J Matthews ◽  
Z Cichon ◽  
W K Aulitzky ◽  
C Y Cheng ◽  
...  
Keyword(s):  
Gene Expression ◽  
Renal Function ◽  
Ureteral Obstruction ◽  
Rabbit Kidney ◽  
Sodium Reabsorption ◽  
Mrna Levels ◽  
Mrna Accumulation ◽  
Contralateral Kidney ◽  
Collecting Ducts ◽  
Clusterin Protein

Clusterin, a protein associated with cell death, has been suggested as a marker of renal injury. Correlation of clusterin gene expression with changes in renal function and quantitative measurement of clusterin protein levels after ureteral obstruction have not been previously reported. With unilateral ureteral obstruction in rabbits as the experimental model, the time course of alterations in renal function, clusterin mRNA accumulation, and concentrations of clusterin protein in serum, urine, and renal tissue were investigated. RBF, GFR, and renal concentrating ability (percent sodium reabsorption and urine osmolarity) all decreased (P < 0.05) in the obstructed kidney from control values within 1 day of ureteral obstruction. Clusterin mRNA levels started to rise in the ipsilateral kidney within 12 h of ureteral obstruction and increased up to 10-fold above control levels after 3 days of obstruction. Hybridization histochemistry showed that clusterin mRNA was initially detectable in collecting ducts and distal tubules within 12 h of ureteral obstruction. After 7 days of obstruction, increased accumulation of clusterin mRNA was also detectable in proximal tubular epithelial cells. Clusterin gene expression remained elevated in collecting ducts after 60 days of obstruction. Clusterin expression in the contralateral kidney was increased twofold over control values after 12 h of obstruction. No increase in clusterin mRNA accumulation was detectable after 24 h in the contralateral kidney. Total clusterin protein in the obstructed kidney increased from 0.59 +/- 0.66 (mean +/- 1 SD) to 2.5 +/- 1.3 micrograms after 7 days of ureteral obstruction (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Gene Expression Changes and Anti-proliferative Effect of Noni (Morinda Citrifolia) Fruit Extract Analysed by Real Time-PCR

Molekul ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. 37
Author(s):  
Hermansyah Hermansyah ◽  
Susilawati Susilawati
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Cell Division ◽  
Real Time ◽  
Cell Division Cycle ◽  
Morinda Citrifolia ◽  
Transcriptional Analysis ◽  
Fruit Extract ◽  
Proliferative Effect

To elucidate the anti-proliferative effect of noni (Morinda citrifolia) fruit extract for a Saccharomyces cerevisiae model organism, analysis of gene expression changes related to cell cycle associated with inhibition effect of noni fruit extract was carried out. Anti-proliferative of noni fruit extract was analyzed using gene expression changes of Saccharomyces cerevisiae (strains FY833 and BY4741).  Transcriptional analysis of genes that play a role in cell cycle was conducted by growing cells on YPDAde broth medium containing 1% (w/v) noni fruit extract, and then subjected using quantitative real-time polymerase chain reaction (RT-PCR).  Transcriptional level of genes CDC6 (Cell Division Cycle-6), CDC20 (Cell Division Cycle-20), FAR1 (Factor ARrest-1), FUS3 (FUSsion-3), SIC1 (Substrate/Subunit Inhibitor of Cyclin-dependent protein kinase-1), WHI5 (WHIskey-5), YOX1 (Yeast homeobOX-1) and YHP1 (Yeast Homeo-Protein-1) increased, oppositely genes expression of DBF4 (DumbBell Forming), MCM1 (Mini Chromosome Maintenance-1) and TAH11 (Topo-A Hypersensitive-11) decreased, while the expression level of genes CDC7 (Cell Division Cycle-7), MBP1 (MIul-box Binding Protein-1) and SWI6 (SWItching deficient-6) relatively unchanged. These results indicated that gene expression changes might associate with anti-proliferative effect from noni fruit extract. These gene expressions changes lead to the growth inhibition of S.cerevisiae cell because of cell cycle defect.

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