Biomedical subjects

N D Perkins

Publications and source records attributed to N D Perkins.

At least 19 recordsLinked to original sources

PubMed2006

Post-translational modifications regulating the activity and function of the nuclear factor kappa B pathway.

The diverse cellular and biological functions of the nuclear factor kappa B (NF-kappaB) pathway, together with the catastrophic consequences of its aberrant regulation, demand specific and highly regulated control of its activity. As described in this review, regulation of the NF-kappaB pathway is brought about through multiple post-translational modifications that control the activity of the core components of NF-kappaB signaling: the IkappaB kinase (IKK) complex, the IkappaB proteins and the NF-kappaB subunits themselves. These regulatory modifications, which include phosphorylation, ubiquitination, acetylation, sumoylation and nitrosylation, can vary, depending on the nature of the NF-kappaB-inducing stimulus. Moreover, they frequently have distinct, sometimes antagonistic, functional consequences and the same modification can have different effects depending on the context. Given the important role of NF-kappaB in human health and disease, understanding these pathways will not only provide valuable insights into mechanism and function, but could also lead to new drug targets and the development of diagnostic and prognostic biomarkers for many pathological conditions.

Animals↗

PubMed2006

Good cop, bad cop: the different faces of NF-kappaB.

Complexes formed from the nuclear factor kappaB (NF-kappaB) family of transcription factors are ubiquitously expressed and are induced by a diverse array of stimuli. This results in their becoming activated in a wide variety of different settings. While the functions of NF-kappaB in many of these contexts have been the subject of intense research and are now well established, it is also clear that there is great diversity in the effects and consequences of NF-kappaB activation. NF-kappaB subunits do not necessarily regulate the same genes, in an identical manner, in all of the different circumstances in which they are induced. This review will discuss the different functions of NF-kappaB, the pathways that modulate NF-kappaB subunit activity and, in contrast to its more commonly thought of role as a promoter of cancer cell growth and survival, the ability of NF-kappaB, under some circumstances, to behave as a tumor suppressor.

Carcinogens↗

PubMed2004

Regulation of NF-kappaB by atypical activators and tumour suppressors.

The RelA(p65) NF-kappaB (nuclear factor kappaB) subunit is typically thought of as being antiapoptotic and tumour-promoting. However, in our laboratory, we have discovered that RelA can inhibit, rather than induce, antiapoptotic gene expression when activated by certain chemotherapeutic drugs, UV light or through the action of the ARF tumour suppressor. These observations explain why RelA can sometimes facilitate rather than inhibit apoptosis and also exhibits tumour-suppressor characteristics in vivo. A better understanding of these processes and an ability to analyse RelA function in tumours could lead to improved cancer diagnosis, choice of therapy and, ultimately, development of new drugs.

Animals↗

PubMed2004

Post-translational modification of RelA(p65) NF-kappaB.

Stimulation with diverse agents activates the NF-kappaB (nuclear factor kappaB) transcription factor, affecting inflammatory and immune responses, proliferation, differentiation, apoptosis and tumourigenesis. Determining how NF-kappaB elicits such distinct responses is essential to understanding NF-kappaB function in diseased tissues. Recent developments illustrating that post-translational modification of NF-kappaB subunits influences their nuclear role are discussed. These observations suggest that diagnosis and new therapies based on reprogramming NF-kappaB activity could be more efficient than total NF-kappaB inhibition.

Animals↗

PubMed2001

SNIP1 inhibits NF-kappa B signaling by competing for its binding to the C/H1 domain of CBP/p300 transcriptional co-activators.

SNIP1 is a 396-amino acid nuclear protein shown to be an inhibitor of the TGF-beta signal transduction pathway and to be important in suppressing transcriptional activation dependent on the co-activators CBP and p300. In this report we show that SNIP1 potently inhibits the activity of NF-kappa B, which binds the C/H1 domain of CBP/p300, but does not interfere with the activity of transcription factors such as p53, which bind to other domains of p300, or factors such as VP16, which are independent of these co-activators. Inhibition of NF-kappa B activity is a function of the N-terminal domain of SNIP1 and involves competition of SNIP1 and the NF-kappa B subunit, RelA/p65, for binding to p300, similar to the mechanism of inhibition of Smad signaling by SNIP1. Immunohistochemical staining shows that expression of SNIP1 is strictly regulated in development and that it colocalizes, in certain tissues, with nuclear staining for RelA/p65 and for p300, suggesting that they may regulate NF-kappa B activity in vivo in a spatially and temporally controlled manner. These data led us to suggest that SNIP1 may be an inhibitor of multiple transcriptional pathways that require the C/H1 domain of CBP/p300.

Animals↗

PubMed2001

UV stimulation induces nuclear factor kappaB (NF-kappaB) DNA-binding activity but not transcriptional activation.

The cellular response to DNA-damaging agents is partly mediated by DNA-binding transcription factors such as p53 and nuclear factor kappaB (NF-kappaB). Typically NF-kappaB activation is associated with resistance to apoptosis. Following stimulation with UV light however, NF-kappaB activation has been shown to be required for programmed cell death. To study this effect further and to determine the relationship between NF-kappaB and p53 function, we have examined the effect of UV light on U2OS cells. UV stimulation resulted in the activation of NF-kappaB DNA-binding and the induction of p53. Surprisingly, and in contrast with tumour necrosis factor alpha stimulation, this UV- induced NF-kappaB was transcriptionally inert. These observations suggest a model in which the NF-kappaB switch from an anti-apoptotic to a pro-apoptotic role within the cell results from modulation of its ability to stimulate gene expression, possibly as a result of the ability of p53 to sequester transcriptional co-activator proteins such as p300/CREB (cAMP-response-element-binding protein)-binding protein.

Animals↗

PubMed2000

Inhibition of the RelA(p65) NF-kappaB subunit by Egr-1.

Induction of transcription from the human immunodeficiency virus 1 long terminal repeat by the RelA (p65) NF-kappaB subunit has been shown to be dependent upon an interaction with the zinc finger DNA-binding domain of Sp1. It was unknown, however, whether NF-kappaB could also interact with other zinc finger-containing transcription factors. In this study we demonstrate that the early growth response transcription factor Egr-1, whose DNA-binding domain shares a high degree of homology with that of Sp1, can also interact with RelA in vitro and regulate NF-kappaB transcriptional activity in vivo. Similar to the interaction with Sp1, the Rel homology domain of RelA interacts with the zinc finger domain of Egr-1. Surprisingly, and in contrast to Sp1, Egr-1 specifically represses RelA transcriptional activity through its zinc finger domain. Moreover, the interaction between RelA and the Egr-1 zinc fingers is mutually exclusive with DNA binding suggesting a model in which Egr-1 directly sequesters NF-kappaB from its target promoters. Because Egr-1 is induced by many of the same stimuli that activate NF-kappaB, this novel transcriptional regulatory mechanism has many implications for the involvement of both factors in cellular processes such as apoptosis and the response to stress and infection.

Base Sequence↗

PubMed2000

The Rel/NF-kappa B family: friend and foe.

The members of the Rel/NF-kappa B family of transcription factors form one of the first lines of defense against infectious diseases and cellular stress. These proteins initiate a highly coordinated response in multiple cell types that effectively counteracts the threat to the health of the organism. Conversely, disruption of the regulatory mechanisms that control the specificity and extent of this response, which results in aberrant activation of NF-kappa B, can be one of the primary causes of a wide range of human diseases. Thus, targeting NF-kappa B might lead to the development of new pharmaceutical reagents that could provide novel treatments for many inflammatory diseases and cancer.

Animals↗

PubMed2000

A novel transcriptional repression domain mediates p21(WAF1/CIP1) induction of p300 transactivation.

The transcriptional coactivators p300 and CREB binding protein (CBP) are important regulators of the cell cycle, differentiation, and tumorigenesis. Both p300 and CBP are targeted by viral oncoproteins, are mutated in certain forms of cancer, are phosphorylated in a cell cycle-dependent manner, interact with transcription factors such as p53 and E2F, and can be found complexed with cyclinE-Cdk2 in vivo. Moreover, p300-deficient cells show defects in proliferation. Here we demonstrate that transcriptional activation by both p300 and CBP is stimulated by coexpression of the cyclin-dependent kinase inhibitor p21(WAF/CIP1). Significantly this stimulation is independent of both the inherent histone acetyltransferase (HAT) activity of p300 and CBP and of the previously reported carboxyl-terminal binding site for cyclinE-Cdk2. Rather, we describe a previously uncharacterized transcriptional repression domain (CRD1) within p300. p300 transactivation is stimulated through derepression of CRD1 by p21. Significantly p21 regulation of CRD1 is dependent on the nature of the core promoter. We suggest that CRD1 provides a novel mechanism through which p300 and CBP can switch activities between the promoters of genes that stimulate growth and those that enhance cell cycle arrest.

Animals↗

PubMed1999

Transcriptional cross talk between NF-kappaB and p53.

Many cellular stimuli result in the induction of both the tumor suppressor p53 and NF-kappaB. In contrast to activation of p53, which is associated with the induction of apoptosis, stimulation of NF-kappaB has been shown to promote resistance to programmed cell death. These observations suggest that a regulatory mechanism must exist to integrate these opposing outcomes and coordinate this critical cellular decision-making event. Here we show that both p53 and NF-kappaB inhibit each other's ability to stimulate gene expression and that this process is controlled by the relative levels of each transcription factor. Expression of either wild-type p53 or the RelA(p65) NF-kappaB subunit suppresses stimulation of transcription by the other factor from a reporter plasmid in vivo. Moreover, endogenous, tumor necrosis factor alpha-activated NF-kappaB will inhibit endogenous wild-type p53 transactivation. Following exposure to UV light, however, the converse is observed, with p53 downregulating NF-kappaB-mediated transcriptional activation. Both p53 and RelA(p65) interact with the transcriptional coactivator proteins p300 and CREB-binding protein (CBP), and we demonstrate that these results are consistent with competition for a limiting pool of p300/CBP complexes in vivo. These observations have many implications for regulation of the transcriptional decision-making mechanisms that govern cellular processes such as apoptosis. Furthermore, they suggest a previously unrealized mechanism through which dysregulated NF-kappaB can contribute to tumorigenesis and disease.

Apoptosis↗

PubMed1998

Cell cycle regulation of the transcriptional coactivators p300 and CREB binding protein.

To respond to changes in its environment, the cell utilizes mechanisms that integrate extracellular signals with specific changes in gene expression. To better understand these critical regulatory mechanisms, research has focused, for the most part, on the identification of sequence-specific DNA-binding proteins, such as the nuclear factor kappaB (NF-kappaB) or activator protein 1 (AP-1) families of transcription factors, that interact with the promoter and enhancer elements of genes induced or repressed during cellular activation. More recently, however, it has become apparent that non-DNA-binding transcriptional coactivators, such as p300 and CREB binding protein (CBP), previously thought to function primarily as "bridging" proteins between DNA-bound transcription factors and the basal transcription complex, play a critical regulatory role as integrators of diverse signalling pathways with the selective induction of gene expression. In this commentary, we shall discuss the implications of a particular aspect of this growing and expanding field: how cell cycle regulation of p300 and CBP impacts our understanding of cellular differentiation, the response to DNA damage, and oncogenesis.

Cell Cycle↗

PubMed1998

Requirements for enhanced transgene expression by untranslated sequences from the human cytomegalovirus immediate-early gene.

BACKGROUND: The cytomegalovirus immediate early (CMV IE) promoter has been widely used for heterologous expression. Further enhancements of gene expression from this potent promoter may allow for the development of improved gene transfer strategies. We aimed to determine whether inclusion of the first exon (5' untranslated) and first intron of the CMV IE gene would increase heterologous transgene expression in primary target cells and to determine the sequences required for any observed increases. MATERIALS AND METHODS: Comparisons of reporter gene expression were made following transient transfection of vascular smooth muscle cells (VSMCs) with plasmids containing the first exon and intron from the CMV IE gene or deletional mutations. Comparisons were also made using a heterologous promoter (RSV). RESULTS: Gene expression from the CMV IE promoter was increased 5.7-fold in VSMC with the inclusion of the first exon and intron. Similar increases were seen with other target cells and from the heterologous RSV promoter. This increase was associated with an increase in steady-state mRNA. Deletion analyses demonstrated that the enhancement was dependent on the presence of the 5' portion of the first exon while deletion of large segments within the intron was associated with similar levels of expression compared with the parental plasmid. CONCLUSIONS: Inclusion of the first exon and intron from the CMV IE gene increases expression from the CMV IE promoter. This enhancement is seen with the heterologous RSV promoter and is associated with an increase in steady-state mRNA. Deletion analyses suggest that this enhancement is associated with inclusion of sequences within the 5' portion of the first exon and inclusion of an intron.

Animals↗

PubMed1997

Regulation of NF-kappaB by cyclin-dependent kinases associated with the p300 coactivator.

The nuclear factor kappaB (NF-kappaB) transcription factor is responsive to specific cytokines and stress and is often activated in association with cell damage and growth arrest in eukaryotes. NF-kappaB is a heterodimeric protein, typically composed of 50- and 65-kilodalton subunits of the Rel family, of which RelA(p65) stimulates transcription of diverse genes. Specific cyclin-dependent kinases (CDKs) were found to regulate transcriptional activation by NF-kappaB through interactions with the coactivator p300. The transcriptional activation domain of RelA(p65) interacted with an amino-terminal region of p300 distinct from a carboxyl-terminal region of p300 required for binding to the cyclin E-Cdk2 complex. The CDK inhibitor p21 or a dominant negative Cdk2, which inhibited p300-associated cyclin E-Cdk2 activity, stimulated kappaB-dependent gene expression, which was also enhanced by expression of p300 in the presence of p21. The interaction of NF-kappaB and CDKs through the p300 and CBP coactivators provides a mechanism for the coordination of transcriptional activation with cell cycle progression.

CDC2-CDC28 Kinases↗

PubMed1997

Achieving transcriptional specificity with NF-kappa B.

Nuclear Factor-Kappa B (NF-kappa B) was first identified by Sen and Baltimore (1986, Cell 46, 705-716) as a constitutively active transcription factor binding the kappa light chain immunoglobulin enhancer in B cells. Shortly afterwards, the same researchers found NF-kappa B to be present in other cell types in an inactive cytoplasmic form which upon cellular stimulation could be induced to translocate to the nucleus and bind DNA. Subsequently, it has been demonstrated that NF-kappa B performs a critical role as a regulator of the immune system, the response to stress, apoptosis, viral replication and is involved in many diseases, leading to it becoming one of the most intensively studied transcription factors of the last decade. The pivotal role played by NF-kappa B is illustrated not only by the great diversity of genes that it regulates, but also by the large variety of stimuli leading to its activation. This article will address how NF-kappa B, a ubiquitously expressed transcription factor composed of dimers formed from five subunits, differentially regulates the expression of such a diverse array of genes with different functions, in different cell types and at different times. Recent research indicates that this behavioral diversity arises from a delicately balanced network of protein: protein interactions: NF-kappa B activity is determined not only through its regulated nuclear localization but is also dependent on the cellular context in which it is found.

Animals↗

PubMed1997

Distinct domains of adenovirus E1A interact with specific cellular factors to differentially modulate human immunodeficiency virus transcription.

Transcription of human immunodeficiency virus (HIV) type 1 and other viruses is regulated by the transcription factor NF-kappaB, which interacts with the multifunctional cellular protein p300. p300, originally identified by its ability to bind adenovirus early region 1A (E1A), has also been shown to regulate HIV transcription through its interaction with NF-kappaB. The 13S form of E1A activates HIV gene expression, while the 12S form represses its transcription. In this report, we have investigated whether these divergent effects of E1A are dependent upon common or distinct cellular cofactors, including p300, pRb, and the TATA box-binding protein (TBP). Unlike activation in the absence of E1A, cooperative stimulation of HIV gene expression by 13S E1A and RelA was independent of the ability of E1A to bind p300 but was critically dependent on the E1A CR3 region which associates with TBP. In contrast, inhibition of basal HIV gene expression by the 12S form of E1A was dependent on p300 but not pRb or TBP. Interestingly, mutation of the CR2 region of 12S E1A responsible for pRb binding abolished the repression of HIV transcription stimulated by tumor necrosis factor alpha, suggesting that repression of cytokine-activated transcription involves cofactors different from those used in unstimulated cells. Repression and activation of HIV transcription by different forms of E1A are mediated by distinct sets of cellular cofactors. These findings suggest that E1A has evolved to interact by alternative mechanisms with a transcriptional coactivator and its associated cofactors to differentially modulate cellular and viral gene expression.

Adenovirus E1A Proteins↗

PubMed1996

Role of the p21 cyclin-dependent kinase inhibitor in limiting intimal cell proliferation in response to arterial injury.

Arterial injury induces a series of proliferative, vasoactive, and inflammatory responses that lead to vascular proliferative diseases, including atherosclerosis and restenosis. Although several factors have been defined which stimulate this process in vivo, the role of specific cellular gene products in limiting this response is not well understood. The p21 cyclin-dependent kinase inhibitor affects cell cycle progression, senescence, and differentiation in transformed cells, but its expression in injured blood vessels has not been investigated. In this study, we report that p21 protein is induced in porcine arteries following balloon catheter injury and suggest that p21 is likely to play a role in limiting arterial cell proliferation in vivo. Vascular endothelial and smooth muscle cell growth was arrested through the ability of p21 to inhibit progression through the G1 phase of the cell cycle. Following injury to porcine arteries, p21 gene product was detected in the neointima and correlated inversely with the location and kinetics of intimal cell proliferation. Direct gene transfer of p21 using an adenoviral vector into balloon injured porcine arteries inhibited the development of intimal hyperplasia. Taken together, these findings suggest that p21, and possibly related cyclin-dependent kinase inhibitors, may normally regulate cellular proliferation following arterial injury, and strategies to increase its expression may prove therapeutically beneficial in vascular diseases.

Adenoviridae↗

PubMed1996

A cooperative interaction of human T-cell leukemia virus type 1 Tax with the p21 cyclin-dependent kinase inhibitor activates the human immunodeficiency virus type 1 enhancer.

Interactions between the Tax transactivator of human T-cell leukemia virus type 1 (HTLV-1) and a cell cycle regulatory protein have been examined. We report cooperative stimulation of human immunodeficiency virus type 1 gene expression by Tax and a regulator of cell cycle progression, the p21 cyclin-dependent kinase inhibitor (CKI). This cooperativity results from the effect of p21 on transcriptional coactivation by Tax-induced NF-kappaB. This effect was abrogated by a mutation in Tax which specifically eliminated NF-kappaB induction, was inhibitable by IkappaB-alpha, and was markedly reduced in human immunodeficiency virus reporter plasmids with mutant kappaB sites. These studies demonstrate that transcriptional activation by Tax is influenced by cell cycle regulatory proteins.

Cyclin-Dependent Kinase Inhibitor p21↗

PubMed1995

Cytokine induction of nuclear factor kappa B in cycling and growth-arrested cells. Evidence for cell cycle-independent activation.

Nuclear factor kappa B (NF-kappa B) is a pleiotropic transcription factor which regulates the expression of a large number of cellular and viral genes. Induction of NF-kappa B has been shown previously to occur during cell cycle transition from G0 to G1, but the relationship of cytokine induction of this transcription factor to cell cycling has not been directly addressed. Here we examine the inductions of NF-kappa B in serum-deprived and cycling cells in response to tumor necrosis factor-alpha (TNF-alpha). In 3T3 fibroblasts deprived of serum, and in the temperature-sensitive G2 phase mutant carcinoma line FT210, we find that NF-kappa B DNA binding activity is rapidly induced upon addition of TNF-alpha. In addition, NF-kappa B induction in cycling cells occurs without a significant change in cell cycle distribution. These data reveal that NF-kappa B is rapidly induced by TNF-alpha in both proliferating and arrested cells and suggest that distinct activation pathways can lead to cell cycle-dependent or -independent induction of NF-kappa B.

3T3 Cells↗