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.

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 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.

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.

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 Mutational analysis of the DNA-binding domain of the CYS3 regulatory protein of Neurospora crassa.

1991 ◽  
Vol 11 (9) ◽  
pp. 4356-4362 ◽  
Author(s):  
M N Kanaan ◽  
G A Marzluf
Keyword(s):  
Dna Binding ◽  
Neurospora Crassa ◽  
Mutational Analysis ◽  
Regulatory Gene ◽  
Regulatory Protein ◽  
Binding Activity ◽  
Binding Domain ◽  
Site Directed Mutagenesis ◽  
Dna Binding Domain ◽  
Dna Binding Activity

cys-3, the major sulfur regulatory gene of Neurospora crassa, activates the expression of a set of unlinked structural genes which encode sulfur catabolic-related enzymes during conditions of sulfur limitation. The cys-3 gene encodes a regulatory protein of 236 amino acid residues with a leucine zipper and an upstream basic region (the b-zip region) which together may constitute a DNA-binding domain. The b-zip region was expressed in Escherichia coli to examine its DNA-binding activity. The b-zip domain protein binds to the promoter region of the cys-3 gene itself and of cys-14, the sulfate permease II structural gene. A series of CYS3 mutant proteins obtained by site-directed mutagenesis were expressed and tested for function, dimer formation, and DNA-binding activity. The results demonstrate that the b-zip region of cys-3 is critical for both its function in vivo and specific DNA-binding in vitro.

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 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.

Cooperative DNA binding of the human HoxB5 (Hox-2.1) protein is under redox regulation in vitro

1993 ◽  
Vol 13 (8) ◽  
pp. 4609-4617
Author(s):  
C K Galang ◽  
C A Hauser
Keyword(s):  
Dna Binding ◽  
Redox Regulation ◽  
Mutational Analysis ◽  
Cysteine Residue ◽  
Binding Activity ◽  
Nf Kappa B ◽  
Amino Terminal ◽  
Dna Binding Activity ◽  
Specific Oxidation

The human HoxB5 (Hox-2.1) gene product is a sequence-specific DNA binding protein. Cooperative interactions stabilize in vitro DNA binding of the HoxB5 protein to tandem binding sites by at least 100-fold relative to binding to a single site. The HoxB5 homeodomain is sufficient for sequence-specific DNA binding but not for cooperative DNA binding. Here we report that the additional protein sequence required for cooperativity is a small domain adjacent to the homeodomain on the amino-terminal side. We further show that cooperative DNA binding is under redox regulation. The HoxB5 protein binds to DNA in vitro both when oxidized or reduced but binds cooperatively only when oxidized. Mutational analysis has revealed that the cysteine residue in the turn between homeodomain helices 2 and 3 is necessary for cooperative binding and redox regulation. The enhanced DNA binding of oxidized HoxB5 protein is the opposite of the redox regulation reported for other mammalian transcription factors such as Fos, Jun, USF, NF-kappa B, c-Myb, and v-Rel, in which oxidation of cysteine residues inhibits DNA binding. Thus, specific oxidation of nuclear proteins is a potential regulatory mechanism that can act to either decrease or increase their DNA binding activity.

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