scholarly journals Protein-DNA interactions at the major and minor promoters of the divergently transcribed dhfr and rep3 genes during the Chinese hamster ovary cell cycle.

1996 ◽  
Vol 16 (2) ◽  
pp. 634-647 ◽  
Author(s):  
J Wells ◽  
P Held ◽  
S Illenye ◽  
N H Heintz
Keyword(s):  
Phase Transition ◽  
Cell Cycle ◽  
Dna Binding ◽  
Binding Sites ◽  
Binding Activity ◽  
Dnase I ◽  
S Phase ◽  
Ovary Cell ◽  
Dna Binding Activity ◽  
Genomic Footprinting

In mammals, two TATA-less bidirectional promoters regulate expression of the divergently transcribed dihydrofolate reductase (dhfr) and rep3 genes. In CHOC 400 cells, dhfr mRNA levels increase about fourfold during the G1-to-S phase transition of the cell cycle, whereas the levels of rep3 transcripts vary less than twofold during this time. To assess the role of DNA-binding proteins in transcriptional regulation of the dhfr and rep3 genes, the major and minor dhfr-rep3 promoter regions were analyzed by high-resolution genomic footprinting during the cell cycle. At the major dhfr promoter, prominent DNase I footprints over four upstream Sp1 binding sites did not vary throughout G1 and entry into the S phase. Genomic footprinting revealed that a protein is constitutively bound to the overlapping E2F sites throughout the G1-to-S phase transition, an interaction that is most evident on the transcribed template strand. On the nontranscribed strand, multiple changes in the DNase I cleavage pattern are observed during transit through G1 and entry into the S phase. By using gel mobility shift assays and a series of sequence-specific probes, two different species of E2F were shown to interact with the dhfr promoter during the cell cycle. The DNA binding activity of one E2F species, which preferentially recognizes the sequence TTTGGCGC, did not vary significantly during the cell cycle. The DNA binding activity of the second E2F species, which preferentially recognizes the sequence TTTCGCGC, increased during the G1-to-S phase transition. Together, these results indicate that Sp1 and the species of E2F that binds TTTGGCGC participate in the formation of a basal transcription complex, while the species of E2F that binds TTTCGCGC regulates dhfr gene expression during the G1-to-S phase transition. At the minor promoter, DNase I footprints at a consensus c-Myc binding site and three Sp1 binding sites showed little variation during the G1-to-S phase transition. In addition to protein binding at sequences known to be involved in the regulation of transcription, genomic footprinting of the entire promoter region also showed that a protein factor is constitutively bound to the first intron of the rep3 gene.

Cell cycle regulation of H2b histone octamer DNA-binding activity in Chinese hamster lung fibroblasts

1989 ◽  
Vol 9 (2) ◽  
pp. 869-873
Author(s):  
M Ito ◽  
A Sharma ◽  
A S Lee ◽  
R Maxson
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Mutational Analysis ◽  
Chinese Hamster ◽  
Binding Activity ◽  
S Phase ◽  
Lung Fibroblasts ◽  
Histone Genes ◽  
Dna Binding Activity ◽  
Cycle Dependent

The promoter regions of H2b histone genes contain a 14-base-pair element which includes the octamer ATTTGCAT. Mutational analysis has implicated the octamer element in the cell cycle-dependent expression of H2b histone genes. In this report, we address the question of whether the DNA-binding activity of the octamer transcription factor is itself cell cycle regulated. By using a gel mobility shift assay, we measured the relative amounts of octamer-binding activity during various phases of the cell cycle in serum-synchronized Chinese hamster fibroblasts. We found that the activity increased approximately fivefold between late G1 phase and early S phase and then decreased threefold between late S phase and G2 phase. These cell cycle-dependent changes in octamer DNA-binding activity may in part account for the selective transcription of H2b histone genes in late G1 and S phases.

scholarly journals p53 transactivation and protein accumulation are independently regulated by UV light in different phases of the cell cycle.

1997 ◽  
Vol 17 (6) ◽  
pp. 3074-3080 ◽  
Author(s):  
T Haapajärvi ◽  
K Pitkänen ◽  
M Tsubari ◽  
M Laiho
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Target Genes ◽  
Binding Activity ◽  
S Phase ◽  
Protein Stabilization ◽  
Cycle Phase ◽  
Protein Accumulation ◽  
Dna Binding Activity ◽  
G1 Cells

DNA damage-induced activation of the p53 tumor suppressor gene is suggested to be central in the cellular damage response pathway. In this study, we analyzed the responses of p53 to UVC radiation in synchronized mouse fibroblasts in terms of p53 accumulation, transcriptional activation, and sequence-specific DNA-binding activity. UVC was found to induce accumulation of p53 cell cycle dependently in G1/S- and S-phase cells but not in G0 or G1 cells. In contrast, p53 transcriptional activity and its target genes, p21 and GADD45, were stimulated by UVC in G0 and G1 cells in the absence of detectable p53 protein. The accumulation of p53 and increased p21 and GADD45 expression were replication dependent in S-phase cells. Interestingly, sequence-specific p53 DNA-binding activity was stimulated also replication independently in S phase, though the effect was not conveyed to stimulation of p53 target genes, suggesting that additional events are required for p53-stimulated gene expression. The results show that opposed to the cell cycle dependence of p53 accumulation, the UVC-mediated transactivation by p53 is independent of the cell cycle phase and protein stabilization.

scholarly journals Cell cycle regulation of H2b histone octamer DNA-binding activity in Chinese hamster lung fibroblasts.

1989 ◽  
Vol 9 (2) ◽  
pp. 869-873 ◽  
Author(s):  
M Ito ◽  
A Sharma ◽  
A S Lee ◽  
R Maxson
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Mutational Analysis ◽  
Chinese Hamster ◽  
Binding Activity ◽  
S Phase ◽  
Lung Fibroblasts ◽  
Histone Genes ◽  
Dna Binding Activity ◽  
Cycle Dependent

The promoter regions of H2b histone genes contain a 14-base-pair element which includes the octamer ATTTGCAT. Mutational analysis has implicated the octamer element in the cell cycle-dependent expression of H2b histone genes. In this report, we address the question of whether the DNA-binding activity of the octamer transcription factor is itself cell cycle regulated. By using a gel mobility shift assay, we measured the relative amounts of octamer-binding activity during various phases of the cell cycle in serum-synchronized Chinese hamster fibroblasts. We found that the activity increased approximately fivefold between late G1 phase and early S phase and then decreased threefold between late S phase and G2 phase. These cell cycle-dependent changes in octamer DNA-binding activity may in part account for the selective transcription of H2b histone genes in late G1 and S phases.

Androgen-dependent and DNA binding-independent association of androgen receptor with chromatic regions coding androgen-induced non-coding RNAs

2021 ◽  
Author(s):  
Takahiro Sawada ◽  
Koichi Nishimura ◽  
Jinichi Mori ◽  
Yoshiaki Kanemoto ◽  
Alexander Kouzmenko ◽  
...  
Keyword(s):  
Dna Binding ◽  
Binding Sites ◽  
Specific Antigen ◽  
Binding Activity ◽  
Lncap Cells ◽  
Coding Regions ◽  
Dna Binding Activity ◽  
Genome Wide ◽  
Non Coding Rnas ◽  
Independent Association

Abstract Androgen induces the binding of its receptor (AR) to androgen-responsive elements (AREs), while genome-wide studies showed that most androgen-induced AR binding sites on chromatin were unrelated to AREs. Enhancer RNAs (eRNAs), a class of non-coding RNAs(ncRNAs), are transcribed from super-enhancers (SEs), and trigger the formation of large ribonucleoprotein (RNP) condensates of transcription factors. By in silico search, an SE is found to be located on the locus of KLK3 that encodes prostate specific antigen (PSA). On the KLK3 SE, androgen-induced expression of ncRNAs was detected and designated as KLK3eRNAs in LNCaP cells, and androgen-induced association of AR and FOXA1 on the KLK3eRNA coding regions was detected. Such androgen-induced association of an AR mutant lacking DNA binding activity on the KLK3eRNA coding regions was undetectable on an exogenous ARE. Thus, the present findings suggest a molecular basis of androgen-induced association of AR with chromatin on ARE-unrelated sequences.

Evidence for DNA binding activity of numatrin (B23), a cell cycle-regulated nuclear matrix protein

1990 ◽  
Vol 1087 (2) ◽  
pp. 127-136 ◽  
Author(s):  
Nili Feuerstein ◽  
James J. Mond ◽  
Paul R. Kinchington ◽  
Robert Hickey ◽  
Marja-Liisa Karjalainen Lindsberg ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Nuclear Matrix ◽  
Matrix Protein ◽  
Binding Activity ◽  
Nuclear Matrix Protein ◽  
Dna Binding Activity ◽  
A Cell

scholarly journals Constitutive c-Myb amino-terminal phosphorylation and DNA binding activity uncoupled during entry and passage through the cell cycle

Oncogene ◽  
2001 ◽  
Vol 20 (14) ◽  
pp. 1784-1792 ◽  
Author(s):  
Alina Cures ◽  
Colin House ◽  
Chie Kanei-Ishii ◽  
Bruce Kemp ◽  
Robert G Ramsay
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Binding Activity ◽  
Amino Terminal ◽  
Dna Binding Activity

scholarly journals Phosphorylation Controls Ikaros's Ability To Negatively Regulate the G1-S Transition

2004 ◽  
Vol 24 (7) ◽  
pp. 2797-2807 ◽  
Author(s):  
Pablo Gómez-del Arco ◽  
Kazushige Maki ◽  
Katia Georgopoulos
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Cell Cycle Progression ◽  
Rapid Development ◽  
Binding Activity ◽  
Casein Kinase ◽  
Cycle Progression ◽  
Dna Binding Activity ◽  
Increased Activity ◽  
Inactivating Mutations

ABSTRACT Ikaros is a key regulator of lymphocyte proliferative responses. Inactivating mutations in Ikaros cause antigen-mediated lymphocyte hyperproliferation and the rapid development of leukemia and lymphoma. Here we show that Ikaros's ability to negatively regulate the G1-S transition can be modulated by phosphorylation of a serine/threonine-rich conserved region (p1) in exon 8. Ikaros phosphorylation in p1 is induced during the G1-S transition. Mutations that prevent phosphorylation in p1 increase Ikaros's ability to impede cell cycle progression and its affinity for DNA. Casein kinase II, whose increased activity in lymphocytes leads to transformation, is a key player in Ikaros p1 phosphorylation. We thus propose that Ikaros's activity as a regulator of the G1-S transition is controlled by phosphorylation in response to signaling events that downmodulate its DNA binding activity.

scholarly journals DNA-binding properties of the Drosophila melanogaster zeste gene product.

1988 ◽  
Vol 8 (2) ◽  
pp. 615-623 ◽  
Author(s):  
A Mansukhani ◽  
A Crickmore ◽  
P W Sherwood ◽  
M L Goldberg
Keyword(s):  
Dna Binding ◽  
Dna Sequences ◽  
Binding Sites ◽  
Fusion Proteins ◽  
Genetic Interaction ◽  
Binding Activity ◽  
Transcriptional Unit ◽  
Dna Binding Activity ◽  
Zinc Finger Motifs ◽  
White Locus

The ability of the zeste moiety of beta-galactosidase-zeste fusion proteins synthesized in Escherichia coli to bind specific DNA sequences was examined. Such fusion proteins recognize a region of the white locus upstream of the start of transcription; this region has previously been shown to be required for genetic interaction between the zeste and white loci. Another strong binding site was localized to a region between 50 and 205 nucleotides before the start of the Ubx transcriptional unit; expression of the bithorax complex is also known to be influenced by the zeste locus. Weaker binding sites were also seen in the vicinity of the bxd and Sgs-4 genes, but it is currently unclear whether these binding sites play a role in transvection effects. The DNA-binding activity of the zeste protein is restricted to a domain of approximately 90 amino acids near the N terminus. This domain does not appear to contain homeobox or zinc finger motifs found in other DNA-binding proteins. The DNA-binding domain is not disrupted by any currently characterized zeste mutations.

scholarly journals The evaluation of anoxia responsive E2F DNA binding activity in the red eared slider turtle, Trachemys scripta elegans

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4755 ◽  
Author(s):  
Kyle K. Biggar ◽  
Kenneth B. Storey
Keyword(s):  
Cell Cycle ◽  
Transcription Factor ◽  
Dna Binding ◽  
Complex Dynamics ◽  
Binding Activity ◽  
Trachemys Scripta ◽  
Model Organisms ◽  
Trachemys Scripta Elegans ◽  
Dna Binding Activity

In many cases, the DNA-binding activity of a transcription factor does not change, while its transcriptional activity is greatly influenced by the make-up of bound proteins. In this study, we assessed the protein composition and DNA-binding ability of the E2F transcription factor complex to provide insight into cell cycle control in an anoxia tolerant turtle through the use of a modified ELISA protocol. This modification also permits the use of custom DNA probes that are tailored to a specific DNA binding region, introducing the ability to design capture probes for non-model organisms. Through the use of EMSA and ELISA DNA binding assays, we have successfully determined the in vitro DNA binding activity and complex dynamics of the Rb/E2F cell cycle regulatory mechanisms in an anoxic turtle, Trachemys scripta elegans. Repressive cell cycle proteins (E2F4, Rb, HDAC4 and Suv39H1) were found to significantly increase at E2F DNA-binding sites upon anoxic exposure in anoxic turtle liver. The lack of p130 involvement in the E2F DNA-bound complex indicates that anoxic turtle liver may maintain G1 arrest for the duration of stress survival.

Efficient transcription of the glycolytic gene ADH1 and three translational component genes requires the GCR1 product, which can act through TUF/GRF/RAP binding sites

1990 ◽  
Vol 10 (2) ◽  
pp. 859-862
Author(s):  
G M Santangelo ◽  
J Tornow
Keyword(s):  
Gene Expression ◽  
Saccharomyces Cerevisiae ◽  
Dna Binding ◽  
Binding Sites ◽  
Binding Activity ◽  
Heterologous Gene ◽  
Dna Binding Activity ◽  
Glycolytic Gene

Glycolytic gene expression in Saccharomyces cerevisiae is thought to be activated by the GCR and TUF proteins. We tested the hypothesis that GCR function is mediated by TUF/GRF/RAP binding sites (UASRPG elements). We found that UASRPG-dependent activation of a heterologous gene and transcription of ADH1, TEF1, TEF2, and RP59 were sensitive to GCR1 disruption. GCR is not required for TUF/GRF/RAP expression or in vitro DNA-binding activity.

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