Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 6 de 6
Filter
1.
Oncogene ; 32(45): 5261-71, 2013 Nov 07.
Article in English | MEDLINE | ID: mdl-23318434

ABSTRACT

The tyrosine kinase c-Abl (or Abl) and the prolyl-isomerase Pin1 cooperatively activate the transcription factor p73 by enhancing recruitment of the acetyltransferase p300. As the transcription factor c-Myc (or Myc) is a known target of Pin1 and p300, we hypothesized that it might be regulated in a similar manner. Consistent with this hypothesis, overexpression of Pin1 augmented the interaction of Myc with p300 and transcriptional activity. The action of Abl, however, was more complex than predicted. On one hand, Abl indirectly enhanced phosphorylation of Myc on Ser 62 and Thr 58, its association with Pin1 and p300 and its acetylation by p300. These effects of Abl were exerted through phosphorylation of substrate(s) other than Myc itself. On the other hand, Abl interacted with the C-terminal domain of Myc and phosphorylated up to five tyrosine residues in its N-terminus, the principal of which was Y74. Indirect immunofluorescence or immunohistochemical staining suggested that the Y74-phosphorylated form of Myc (Myc-pY74) localized to the cytoplasm and coexisted either with active Abl in a subset of mammary carcinomas or with Bcr-Abl in chronic myeloid leukemia. In all instances, Myc-pY74 constituted a minor fraction of the cellular Myc protein. Thus, our data unravel two potential effects of Abl on Myc: first, Abl signaling can indirectly augment acetylation of Myc by p300, and most likely also its transcriptional activity in the nucleus; second, Abl can directly phosphorylate Myc on tyrosine: the resulting form of Myc appears to be cytoplasmic, and its presence correlates with Abl activation in cancer.


Subject(s)
Breast Neoplasms/metabolism , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Proto-Oncogene Proteins c-abl/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Acetylation , Animals , Breast Neoplasms/genetics , Cell Line, Tumor , E1A-Associated p300 Protein/metabolism , Fusion Proteins, bcr-abl/metabolism , HEK293 Cells , HeLa Cells , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Mice , NIMA-Interacting Peptidylprolyl Isomerase , Peptidylprolyl Isomerase/metabolism , Phosphorylation , Proto-Oncogene Proteins c-abl/genetics , Proto-Oncogene Proteins c-myc/genetics , RNA Interference , RNA, Small Interfering , Signal Transduction
2.
Oncogene ; 31(13): 1695-709, 2012 Mar 29.
Article in English | MEDLINE | ID: mdl-21860422

ABSTRACT

The transition from quiescence to proliferation is a key regulatory step that can be induced by serum stimulation in cultured fibroblasts. The transcription factor Myc is directly induced by serum mitogens and drives a secondary gene expression program that remains largely unknown. Using mRNA profiling, we identify close to 300 Myc-dependent serum response (MDSR) genes, which are induced by serum in a Myc-dependent manner in mouse fibroblasts. Mapping of genomic Myc-binding sites by ChIP-seq technology revealed that most MDSR genes were directly targeted by Myc, but represented a minor fraction (5.5%) of all Myc-bound promoters (which were 22.4% of all promoters). Other target loci were either induced by serum in a Myc-independent manner, were not significantly regulated or were negatively regulated. MDSR gene products were involved in a variety of processes, including nucleotide biosynthesis, ribosome biogenesis, DNA replication and RNA control. Of the 29 MDSR genes targeted by RNA interference, three showed a requirement for cell-cycle entry upon serum stimulation and 11 for long-term proliferation and/or survival. Hence, proper coordination of key regulatory and biosynthetic pathways following mitogenic stimulation relies upon the concerted regulation of multiple Myc-dependent genes.


Subject(s)
Chromosome Mapping , Fibroblasts/metabolism , Gene Expression Regulation , Proto-Oncogene Proteins c-myc/metabolism , Serum/physiology , Animals , Cell Line , Mice
3.
Oncogene ; 28(3): 422-30, 2009 Jan 22.
Article in English | MEDLINE | ID: mdl-18978814

ABSTRACT

Myc and transforming growth factor-beta (TGFbeta) signaling are mutually antagonistic, that is Myc suppresses the activation of TGFbeta-induced genes, whereas TGFbeta represses c-myc transcription. Here, we report a positive role for Myc in the TGFbeta response, consisting in the induction of an epithelial-to-mesenchymal transition (EMT) and the activation of the EMT-associated gene Snail. Knockdown of either Myc or the TGFbeta effectors SMAD3/4 in epithelial cells eliminated Snail induction by TGFbeta. Both Myc and SMAD complexes targeted the Snail promoter in vivo, DNA binding occurring in a mutually independent manner. Myc was bound prior to TGFbeta treatment, and was required for rapid Snail activation upon SMAD binding induced by TGFbeta. On the other hand, c-myc downregulation by TGFbeta was a slower event, occurring after Snail induction. The response of Snail to another cytokine, hepatocyte growth factor (HGF), also depended on Myc and SMAD4. Thus, contrary to their antagonistic effects on Cip1 and INK4b, Myc and SMADs cooperate in signal-dependent activation of Snail in epithelial cells. Although Myc also targeted the Snail promoter in serum-stimulated fibroblasts, it was dispensable for its activation in these conditions, further illustrating that the action of Myc in transcriptional regulation is context-dependent. Our findings suggest that Myc and TGFbeta signaling may cooperate in promoting EMT and metastasis in carcinomas.


Subject(s)
Epithelial Cells/metabolism , Mesoderm/metabolism , Proto-Oncogene Proteins c-myc/physiology , Transcription Factors/genetics , Transforming Growth Factor beta/metabolism , Animals , Cells, Cultured , Chromatin Immunoprecipitation , Humans , Immunoblotting , Mammary Glands, Animal/cytology , Mammary Glands, Animal/metabolism , Mesoderm/cytology , Mice , RNA, Small Interfering/pharmacology , Smad2 Protein/antagonists & inhibitors , Smad2 Protein/genetics , Smad2 Protein/metabolism , Smad3 Protein/antagonists & inhibitors , Smad3 Protein/genetics , Smad3 Protein/metabolism , Smad4 Protein/antagonists & inhibitors , Smad4 Protein/genetics , Smad4 Protein/metabolism , Snail Family Transcription Factors , Transcription Factors/metabolism , Transcription, Genetic , Transforming Growth Factor beta/genetics
4.
Bone Marrow Transplant ; 32(8): 791-3, 2003 Oct.
Article in English | MEDLINE | ID: mdl-14520423

ABSTRACT

Allogeneic stem cell transplantation (allo-SCT) is an effective and potentially curative treatment for some cases of multiple myeloma (MM). The curative efficacy of allo-SCT may be largely attributed to its immunological activity, the graft-versus-myeloma (GVM) effect. To evaluate the kinetics of residual myeloma cells, we analyzed the follow-up bone marrow samples of three MM patients by means of a real-time molecular assay. We identified a consistent correlation between onset of graft-versus-host disease and disease response. These data suggest that real-time molecular follow-up can be used to monitor the GVM effect and that it can be employed in the clinical setting to tailor post transplant immunomodulation.


Subject(s)
Genetic Testing/methods , Graft vs Tumor Effect , Hematopoietic Stem Cell Transplantation/adverse effects , Multiple Myeloma/therapy , Neoplasm, Residual/diagnosis , Adult , Bone Marrow Transplantation/adverse effects , Follow-Up Studies , Genetic Markers , Humans , Male , Middle Aged , Neoplasm, Residual/genetics , Polymerase Chain Reaction/methods
5.
Nat Med ; 7(11): 1232-5, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11689888

ABSTRACT

HIV-1 infects target cells via a receptor complex formed by CD4 and a chemokine receptor, primarily CCR5 or CXCR4 (ref. 1). Commonly, HIV-1 transmission is mediated by CCR5-tropic variants, also designated slow/low, non-syncytia-inducer or macrophage-tropic, which dominate the early stages of HIV-1 infection and frequently persist during the entire course of the disease. In contrast, HIV-1 variants that use CXCR4 are typically detected at the later stages, and are associated with a rapid decline in CD4+ T cells and progression to AIDS (refs. 2,7-11). Disease progression is also associated with the emergence of concurrent infections that may affect the course of HIV disease by unknown mechanisms. A lymphotropic agent frequently reactivated in HIV-infected patients is human herpesvirus 6 (HHV-6), which has been proposed as a cofactor in AIDS progression. Here we show that in human lymphoid tissue ex vivo, HHV-6 affects HIV-1 infection in a coreceptor-dependent manner, suppressing CCR5-tropic but not CXCR4-tropic HIV-1 replication, as shown with both uncloned viral isolates and isogenic molecular chimeras. Furthermore, we demonstrate that HHV-6 increases the production of the CCR5 ligand RANTES ('regulated upon activation, normal T-cell expressed and secreted'), the most potent HIV-inhibitory CC chemokine, and that exogenous RANTES mimics the effects of HHV-6 on HIV-1, providing a mechanism for the selective blockade of CCR5-tropic HIV-1. Our data suggest that HHV-6 may profoundly influence the course of HIV-1 infection.


Subject(s)
HIV-1/physiology , HIV-1/pathogenicity , Herpesvirus 6, Human/physiology , Chemokine CCL5/biosynthesis , Chemokine CCL5/pharmacology , Culture Techniques , HIV Infections/complications , HIV Infections/etiology , HIV Infections/virology , HIV-1/drug effects , HIV-1/genetics , Humans , Lymphoid Tissue/immunology , Lymphoid Tissue/virology , Receptors, CCR5/physiology , Receptors, CXCR4/physiology , Roseolovirus Infections/complications , Roseolovirus Infections/etiology , Roseolovirus Infections/virology , Virus Replication/drug effects
6.
J Clin Microbiol ; 39(6): 2308-10, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11376078

ABSTRACT

We developed reverse transcriptase (RT) PCR assays for the detection of mRNA from three spliced genes of human herpesvirus 6 (HHV-6), the immediate-early genes U16/U17 and U89/U90 and the late gene U60/U66. Sequence analysis determined the splicing sites of these genes. The new assays may be instrumental in investigating the association between HHV-6 and disease.


Subject(s)
Herpesviridae Infections/diagnosis , Herpesvirus 6, Human/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Genes, Immediate-Early , Genes, Viral , Herpesviridae Infections/virology , Herpesvirus 6, Human/genetics , Humans , Immediate-Early Proteins/genetics , Molecular Sequence Data , RNA Splicing , RNA, Messenger/metabolism , Sequence Analysis, DNA , Viral Proteins/genetics
SELECTION OF CITATIONS
SEARCH DETAIL
...