Transcriptional repression of the E2-containing promoters EIIaE, c-myc, and RB1 by the product of the RB1 gene

1992 ◽  
Vol 12 (8) ◽  
pp. 3431-3438
Author(s):  
P A Hamel ◽  
R M Gill ◽  
R A Phillips ◽  
B L Gallie
Keyword(s):  
Cell Cycle ◽  
Transcriptional Repression ◽  
Susceptibility Gene ◽  
Protein Product ◽  
T Antigen ◽  
Binding Motif ◽  
Regulation Of Transcription ◽  
Expression Of Genes ◽  
Cellular Genes ◽  
A Cell

The protein product of the retinoblastoma susceptibility gene, p110RB1, is a nuclear phosphoprotein [W.H. Lee, J.Y. Shew, F.D. Hong, T.W. Sery, L.A. Donoso, L.J. Young, R. Bookstein, and E.Y. Lee, Nature (London) 329:642-645, 1987] with properties of a cell cycle regulator (K. Buchkovich, L.A. Duffy, and E. Harlow, Cell 58:1097-1105, 1989; P.L. Chen, P. Scully, J.Y. Shew, J.Y. Wang, and W.H. Lee, Cell 58:1193-1198, 1989; J.A. DeCaprio, J.W. Ludlow, D. Lynch, Y. Furukawa, J. Griffin, H. Piwnica-Worms, C.M. Huang, and D.M. Livingston, Cell 58:1085-1095, 1989; and K. Mihara, X.R. Cao, A. Yen, S. Chandler, B. Driscoll, A.L. Murphree, A. TAng, and Y.K. Fung, Science 246:1300-1303, 1989). Although the mechanism of action of p110RB1 remains unknown, several lines of evidence suggest that it plays a role in the regulation of transcription. We now show that overexpression of p110RB1 causes repression of the adenovirus early promoter EIIaE and the promoters of two cellular genes, c-myc and RB1, both of which contain E2F-binding motifs. Mutation of the E2 element in the c-myc promoter abolishes p110RB1 repression. We also demonstrate that a p110RB1 mutant, which is refractory to cell cycle phosphorylation but intact in E1a/large T antigen-binding properties, represses EIIaE with 50- to 80-fold greater efficiency than wild-type p110RB1. These data provide evidence that hypophosphorylated p110RB1 actively represses expression of genes with promoters containing the E2F-binding motif (E2 element).

scholarly journals Transcriptional repression of the E2-containing promoters EIIaE, c-myc, and RB1 by the product of the RB1 gene.

1992 ◽  
Vol 12 (8) ◽  
pp. 3431-3438 ◽  
Author(s):  
P A Hamel ◽  
R M Gill ◽  
R A Phillips ◽  
B L Gallie
Keyword(s):  
Cell Cycle ◽  
Transcriptional Repression ◽  
Susceptibility Gene ◽  
Protein Product ◽  
T Antigen ◽  
Binding Motif ◽  
Regulation Of Transcription ◽  
Expression Of Genes ◽  
Cellular Genes ◽  
A Cell

The protein product of the retinoblastoma susceptibility gene, p110RB1, is a nuclear phosphoprotein [W.H. Lee, J.Y. Shew, F.D. Hong, T.W. Sery, L.A. Donoso, L.J. Young, R. Bookstein, and E.Y. Lee, Nature (London) 329:642-645, 1987] with properties of a cell cycle regulator (K. Buchkovich, L.A. Duffy, and E. Harlow, Cell 58:1097-1105, 1989; P.L. Chen, P. Scully, J.Y. Shew, J.Y. Wang, and W.H. Lee, Cell 58:1193-1198, 1989; J.A. DeCaprio, J.W. Ludlow, D. Lynch, Y. Furukawa, J. Griffin, H. Piwnica-Worms, C.M. Huang, and D.M. Livingston, Cell 58:1085-1095, 1989; and K. Mihara, X.R. Cao, A. Yen, S. Chandler, B. Driscoll, A.L. Murphree, A. TAng, and Y.K. Fung, Science 246:1300-1303, 1989). Although the mechanism of action of p110RB1 remains unknown, several lines of evidence suggest that it plays a role in the regulation of transcription. We now show that overexpression of p110RB1 causes repression of the adenovirus early promoter EIIaE and the promoters of two cellular genes, c-myc and RB1, both of which contain E2F-binding motifs. Mutation of the E2 element in the c-myc promoter abolishes p110RB1 repression. We also demonstrate that a p110RB1 mutant, which is refractory to cell cycle phosphorylation but intact in E1a/large T antigen-binding properties, represses EIIaE with 50- to 80-fold greater efficiency than wild-type p110RB1. These data provide evidence that hypophosphorylated p110RB1 actively represses expression of genes with promoters containing the E2F-binding motif (E2 element).

Gene Expression Profiling in AML and MDS Identifies Genes Located on 5q Which Are Consistently Lower Expressed in Cases with 5q Deletion as Compared to Cases with Normal Karyotype.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 549-549 ◽  
Author(s):  
Claudia Schoch ◽  
Alexander Kohlmann ◽  
Wolfgang Kern ◽  
Sylvia Merk ◽  
Wolfgang Hiddemann ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Signal Transduction ◽  
Normal Karyotype ◽  
Differentially Expressed ◽  
Regulation Of Transcription ◽  
Chromosome 5 ◽  
Expression Of Genes ◽  
Lower Expression ◽  
5Q Deletion

Abstract Deletions of the long arm of chromosome 5 occur either as the sole karyotype abnormality in MDS and AML or as part of a complex aberrant karyotype. It was the aim of this study to analyze the impact of the 5q deletion on the expression levels of genes located on chromosome 5q in AML and MDS. Therefore, gene expression analysis was performed in 344 AML and MDS cases using Affymetrix U133A+B oligonucleotide microarrays. The following subgroups were analyzed: AML with sole 5q deletion (n=7), AML with complex aberrant karyotype (n=83), MDS with sole 5q deletion (n=9), and MDS with complex aberrant karyotype (n=9). These were compared to 200 AML and 36 MDS with normal karyotype. In total, 1313 probe sets representing 603 genes cover sequences located on the long arm of chromosome 5. Overall a significant lower mean expression of all genes located on the long arm of chromosome 5 was observed in subgroups with 5q deletion in comparison to their respective control groups (for all comparisons, p<0.05). 36 genes showed a significantly lower expression in all comparisons. These genes are involved in a variety of different biological processes such as signal transduction (CSNK1A1, DAMS), cell cycle regulation (HDAC3, PFDN1) and regulation of transcription (CNOT8). In addition we performed class prediction using support vector machines (SVM). In one approach all 6 different subgroups were analyzed as one class each. While AML and MDS with normal karyotype as well as AML with complex aberrant karyotype were correctly predicted with high accuracies (97%, 81%, and 92%, respectively) AML and MDS with 5q- sole and MDS with complex aberrant karyotype were frequently misclassified as AML with complex aberrant karyotype. In a second approach only two classes were defined: all cases with 5q deletion combined vs. all cases without 5q deletion. 102 out of 108 cases (94%) with 5q deletion were identified correctly supporting the fact that a distinct gene expression pattern is associated with 5q deletion in general. Performing SVM only with genes located on the long arm of chromosome 5 also resulted in a correct prediction of 92 of 108 (85%) stressing the importance of the expression of genes located on chromosome 5 for these AML and MDS subtypes. The top 100 differentially expressed probe sets between cases with and without 5q deletion represented 74 different annotated genes of which 23 are located on the long arm of chromosome 5. They are involved in a variety of different biological functions such as DNA repair (POLE, RAD21, RAD23B), regulation of transcription (ZNF75A, AF020591, MLLT3, HOXB6), protein biosynthesis (UPF2, TINP1, RPL12, RPL14, RPL15) cell cycle control (GMNN, CSPG6, PFDN1) and signal transduction (HINT1, STK24, APP, CAMLG). 10 of the top 74 genes associated with 5q deletion were involved in the CMYC-pathway with upregulation of RAD21, RAD23B, GMMN, CSPG6, APP, POLE STK24 and STAG2, and downregulation of ACTA2, and RPL12. Ten other genes out of the 74 top differentially expressed genes were involved in the TP53 pathway with upregulation of H1F0, PTPN11 and TAF2 and downregulation of DF, UBE2D2, EEF1A1, IGBP1, PPP2CA, EIF2S3, and NACA. In conclusion, loss of parts of the long arm of chromosome 5 leads to a lower expression of genes located on the long arm of chromosome 5. A specific pattern of functionally related genes was identified which shows a lower expression in AML and MDS subtypes with 5q deletion.

scholarly journals The Human Cytomegalovirus UL82 Gene Product (pp71) Accelerates Progression through the G1 Phase of the Cell Cycle

2003 ◽  
Vol 77 (6) ◽  
pp. 3451-3459 ◽  
Author(s):  
Robert F. Kalejta ◽  
Thomas Shenk
Keyword(s):  
Cell Cycle ◽  
Human Cytomegalovirus ◽  
Cell Cycle Progression ◽  
Protein Product ◽  
S Phase ◽  
Quiescent Cells ◽  
A Cell ◽  
Infected Cells ◽  
The One ◽  
Infectious Cycle

ABSTRACT As viruses are reliant upon their host cell to serve as proper environments for their replication, many have evolved mechanisms to alter intracellular conditions to suit their own needs. For example, human cytomegalovirus induces quiescent cells to enter the cell cycle and then arrests them in late G1, before they enter the S phase, a cell cycle compartment that is presumably favorable for viral replication. Here we show that the protein product of the human cytomegalovirus UL82 gene, pp71, can accelerate the movement of cells through the G1 phase of the cell cycle. This activity would help infected cells reach the late G1 arrest point sooner and thus may stimulate the infectious cycle. pp71 also induces DNA synthesis in quiescent cells, but a pp71 mutant protein that is unable to induce quiescent cells to enter the cell cycle still retains the ability to accelerate the G1 phase. Thus, the mechanism through which pp71 accelerates G1 cell cycle progression appears to be distinct from the one that it employs to induce quiescent cells to exit G0 and subsequently enter the S phase.

scholarly journals Subunit composition determines E2F DNA-binding site specificity.

1997 ◽  
Vol 17 (12) ◽  
pp. 6994-7007 ◽  
Author(s):  
Y Tao ◽  
R F Kassatly ◽  
W D Cress ◽  
J M Horowitz
Keyword(s):  
Cell Cycle ◽  
Dna Binding ◽  
Binding Sites ◽  
Dna Bending ◽  
Susceptibility Gene ◽  
Specific Sequence ◽  
Cellular Genes ◽  
Cycle Dependent

The product of the retinoblastoma (Rb) susceptibility gene, Rb-1, regulates the activity of a wide variety of transcription factors, such as E2F, in a cell cycle-dependent fashion. E2F is a heterodimeric transcription factor composed of two subunits each encoded by one of two related gene families, denoted E2F and DP. Five E2F genes, E2F-1 through E2F-5, and two DP genes, DP-1 and DP-2, have been isolated from mammals, and heterodimeric complexes of these proteins are expressed in most, if not all, vertebrate cells. It is not yet clear whether E2F/DP complexes regulate overlapping and/or specific cellular genes. Moreover, little is known about whether Rb regulates all or a subset of E2F-dependent genes. Using recombinant E2F, DP, and Rb proteins prepared in baculovirus-infected cells and a repetitive immunoprecipitation-PCR procedure (CASTing), we have identified consensus DNA-binding sites for E2F-1/DP-1, E2F-1/DP-2, E2F-4/DP-1, and E2F-4/DP-2 complexes as well as an Rb/E2F-1/DP-1 trimeric complex. Our data indicate that (i) E2F, DP, and Rb proteins each influence the selection of E2F-binding sites; (ii) E2F sites differ with respect to their intrinsic DNA-bending properties; (iii) E2F/DP complexes induce distinct degrees of DNA bending; and (iv) complex-specific E2F sites selected in vitro function distinctly as regulators of cell cycle-dependent transcription in vivo. These data indicate that the specific sequence of an E2F site may determine its role in transcriptional regulation and suggest that Rb/E2F complexes may regulate subsets of E2F-dependent cellular genes.

scholarly journals Polyomavirus Large T Antigen Induces Alterations in Cytoplasmic Signalling Pathways Involving Shc Activation

1999 ◽  
Vol 73 (2) ◽  
pp. 1427-1437 ◽  
Author(s):  
Vanesa Gottifredi ◽  
Giuliana Pelicci ◽  
Eliana Munarriz ◽  
Rossella Maione ◽  
Pier Giuseppe Pelicci ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Growth Factors ◽  
Growth Factor ◽  
Cell Cycle Progression ◽  
Susceptibility Gene ◽  
T Antigen ◽  
Adapter Protein ◽  
Large T Antigen ◽  
Cycle Progression ◽  
Kinase Substrate

ABSTRACT It has been extensively demonstrated that growth factors play a key role in the regulation of proliferation. Several lines of evidence support the hypothesis that for the induction of cell cycle progression in the absence of exogenous growth factors, oncogenes must either induce autocrine growth factor secretion or, alternatively, activate their receptors or their receptor substrates. Cells expressing polyomavirus large T antigen (PyLT) display reduced growth factor requirements, but the mechanisms underlying this phenomenon have yet to be explored. We conducted tests to see whether the reduction in growth factor requirements induced by PyLT was related to alterations of growth factor-dependent signals. To this end, we analyzed the phosphorylation status of a universal tyrosine kinase substrate, the transforming Shc adapter protein, in fibroblasts expressing the viral oncogene. We report that the level of Shc phosphorylation does not decrease in PyLT-expressing fibroblasts after growth factor withdrawal and that this PyLT-mediated effect does not require interaction with protein encoded by the retinoblastoma susceptibility gene. We also found that the chronic activation of the adapter protein is correlated with the binding of Shc to Grb-2 and with defects in the downregulation of mitogen-activated protein kinases. In fibroblasts expressing the nuclear oncoprotein, we also observed the formation of a PyLT-Shc complex that might be involved in constitutive phosphorylation of the adapter protein. Viewed comprehensively, these results suggest that the cell cycle progression induced by PyLT may depend not only on the direct inactivation of nuclear antioncogene products but also on the indirect induction, through the alteration of cytoplasmic pathways, of growth factor-dependent nuclear signals.

scholarly journals Mutagenesis of the pRB Pocket Reveals that Cell Cycle Arrest Functions Are Separable from Binding to Viral Oncoproteins

2000 ◽  
Vol 20 (10) ◽  
pp. 3715-3727 ◽  
Author(s):  
Frederick A. Dick ◽  
Elizabeth Sailhamer ◽  
Nicholas J. Dyson
Keyword(s):  
Cell Cycle ◽  
Binding Site ◽  
Cell Cycle Arrest ◽  
Binding Proteins ◽  
Tumor Cell Line ◽  
T Antigen ◽  
Viral Oncoproteins ◽  
Adenovirus E1a ◽  
Cycle Arrest ◽  
A Cell

ABSTRACT The pocket domain of pRB is required for pRB to arrest the cell cycle. This domain was originally defined as the region of the protein that is necessary and sufficient for pRB's interaction with adenovirus E1A and simian virus s40 large T antigen. These oncoproteins, and other pRB-binding proteins that are encoded by a variety of plant and animal viruses, use a conserved LXCXE motif to interact with pRB. Similar sequences have been identified in multiple cellular pRB-binding proteins, suggesting that the viruses have evolved to target a highly conserved binding site of pRB that is critical for its function. Here we have constructed a panel of pRB mutants in which conserved amino acids that are predicted to make close contacts with an LXCXE peptide were altered. Despite the conservation of the LXCXE binding site throughout evolution, pRB mutants that lack this site are able to induce a cell cycle arrest in a pRB-deficient tumor cell line. This G1 arrest is overcome by cyclin D-cdk4 complexes but is resistant to inactivation by E7. Consequently, mutants lacking the LXCXE binding site were able to induce a G1 arrest in HeLa cells despite the expression of HPV-18 E7. pRB mutants lacking the LXCXE binding site are defective in binding to adenovirus E1A and human papillomavirus type 16 E7 protein but exhibit wild-type binding to E2F or DP, and they retain the ability to interact with CtIP and HDAC1, two transcriptional corepressors that contain LXCXE-like sequences. Consistent with these observations, the pRB mutants are able to actively repress transcription. These observations suggest that viral oncoproteins depend on the LXCXE-binding site of pRB for interaction to a far greater extent than cellular proteins that are critical for cell cycle arrest or transcriptional repression. Mutation of this binding site allows pRB to function as a cell cycle regulator while being resistant to inactivation by viral oncoproteins.

scholarly journals Evidence for a new kind of regulatory gene controlling expression of genes for morphogenesis during the cell cycle in Ustilago violacea

1975 ◽  
Vol 25 (3) ◽  
pp. 253-266 ◽  
Author(s):  
A. W. Day ◽  
J. E. Cummins
Keyword(s):  
Cell Cycle ◽  
Mating Type ◽  
Cell Cycle Control ◽  
Regulatory Gene ◽  
Type Locus ◽  
Ustilago Violacea ◽  
Temporal Properties ◽  
Expression Of Genes ◽  
Separate Control ◽  
A Cell

SUMMARYThe first part of the paper provides strong supportive evidence for the previous findings (Cummins & Day, 1973; Day & Cummins, 1973) that the two alleles of the mating-type locus of the basidiomycete Ustilago violacea have different periods of inducibility during a cell cycle, and that the cell cycle characteristics of each allele are maintained in freshly isolated diploids. This difference in temporal properties of the alleles appears to be the basis of the dominance of allele a2 as it is inducible during a phase of the cell cycle when allele a1 is non-inducible. During G1 both alleles appear to be inducible and apparently ‘neutralize’ each other so that the cell cannot mate.The second part of the paper provides evidence for a unique genetic control mechanism. The evidence suggests that the period of cell cycle inducibility of a locus governing a morphogenetic pathway may be regulated by a separate control gene the cc locus, with two known alleles ccstr(a stringent or restricted period of inducibility) and ccrel (a relaxed or non-restricted period of inducibility). This hypothesis stems from analysis of a diploid that was a1· ccstr/a2· ccrel and showed dominance of allele a2 during the S and G2 phases when freshly isolated, but which became incapable of mating after a period of subculturing. Analysis of haploids derived from this diploid strain showed that both mating-type alleles were functional but that it was now homozygous for ccstr, i.e. of genotype a1· ccstr/a2·ccstr· Thus the temporal and functional aspects of the mating type alleles are determined by different loci. It is postulated that cell cycle control loci may be widespread and serve to regulate the action of genes concerned with morphogenesis in relation to other cell cycle events.

scholarly journals The retinoblastoma protein physically associates with the human cdc2 kinase.

1992 ◽  
Vol 12 (3) ◽  
pp. 971-980 ◽  
Author(s):  
Q J Hu ◽  
J A Lees ◽  
K J Buchkovich ◽  
E Harlow
Keyword(s):  
Cell Cycle ◽  
Kinase Activity ◽  
Susceptibility Gene ◽  
Biochemical Properties ◽  
Protein Product ◽  
S Phase ◽  
Negative Regulator ◽  
Cdc2 Kinase ◽  
Related Enzyme

The protein product (pRB) of the retinoblastoma susceptibility gene functions as a negative regulator of cell proliferation, and its activity appears to be modulated by phosphorylation. Using a new panel of anti-human pRB monoclonal antibodies, we have investigated the biochemical properties of this protein. These antibodies have allowed us to detect a pRB-associated kinase that has been identified as the cell cycle-regulating kinase p34cdc2 or a closely related enzyme. Since this associated kinase phosphorylates pRB at most of the sites used in vivo, these results suggest that this kinase is one of the major regulators of pRB. The associated kinase activity follows the pattern of phosphorylation seen for pRB in vivo. The associated kinase activity is not seen in the G1 phase but appears in the S phase, and the levels continue to increase throughout the remainder of the cell cycle.

The product of the retinoblastoma susceptibility gene has properties of a cell cycle regulatory element

Cell ◽  
1989 ◽  
Vol 58 (6) ◽  
pp. 1085-1095 ◽  
Author(s):  
James A. DeCaprio ◽  
John W. Ludlow ◽  
Dennis Lynch ◽  
Yusuke Furukawa ◽  
James Griffin ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Regulatory Element ◽  
Susceptibility Gene ◽  
A Cell

scholarly journals Cell cycle corruption in a preleukemic ETV6-RUNX1 model exposes RUNX1 addiction as a therapeutic target in acute lymphoblastic leukemia

2020 ◽  
Author(s):  
Jason P Wray ◽  
Elitza M Deltcheva ◽  
Charlotta Boiers ◽  
Simon E Richardson ◽  
Jyoti Bikram Chettri ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Acute Lymphoblastic Leukemia ◽  
Chromatin Immunoprecipitation ◽  
Transcriptional Repression ◽  
Lymphoblastic Leukemia ◽  
Rna Seq ◽  
Chromatin Immunoprecipitation Sequencing ◽  
A Cell ◽  
Cycle Regulation ◽  
Overt Leukemia

SummaryETV6-RUNX1 is the most common translocation in pediatric B-acute lymphoblastic leukemia (B-ALL), shown to arise in utero and to initiate a clinically silent pre-leukemic state. Here, we integrate chromatin immunoprecipitation sequencing (ChIP-seq) and RNA-sequencing (RNA-seq) to characterize the authentic ETV6-RUNX1 regulome and explore its interplay with the native RUNX1 as a substrate for preleukemia. We show that preleukemic initiation is primarily mediated through competition for RUNX1 binding sites and transcriptional repression of the RUNX1 program. Using mass cytometry we functionally demonstrate that as a “first-hit” ETV6-RUNX1 induces a cell cycle impairment in a human pluripotent stem cell (hPSC) model, antagonizing RUNX1-mediated cell cycle regulation. Strikingly, in overt leukemia, genetic perturbation of native RUNX1 protein or its co-factor CBFβ leads to cell death of pediatric and adult B-ALL cells. We demonstrate that RUNX1 addiction can be therapeutically exploited by pharmacological inhibition using an allosteric CBFβ inhibitor.

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