HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 275, Issue 44, 34021-34024, November 3, 2000

This Article Abstract Full Text (PDF) All Versions of this Article: 275/44/34021    most recent C000507200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Foehr, E. D. Articles by Greene, W. C. Search for Related Content PubMed PubMed Citation Articles by Foehr, E. D. Articles by Greene, W. C. Social Bookmarking                      What's this?

ACCELERATED PUBLICATION
The NF-B-inducing Kinase Induces PC12 Cell Differentiation and Prevents Apoptosis^*

Erik D. Foehr^§, Jan Bohuslav, Lin-Feng Chen, Carlos DeNoronha, Romas Geleziunas, Xin Lin, Alison O'Mahony, and Warner C. Greene^¶

From the  Gladstone Institute of Virology and Immunology and the Departments of ^¶ Medicine and  Microbiology and Immunology, University of California, San Francisco, California 94141

Received for publication, July 31, 2000

	ABSTRACT

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

NF-B has been implicated in the survival and differentiation of PC12 cells. In this study, we examined the effect of the NF-B-inducing kinase (NIK) on these processes. When inducibly expressed in PC12 cells, a kinase-proficient but not -deficient form of NIK promoted neurite process formation and mediated anti-apoptotic signaling. As expected, NIK expression led to IB kinase activation and induced nuclear translocation of NF-B. However, NIK-induced neurite outgrowth was only partially blocked by concomitant expression of a nondegradable form of IB that completely blocks NF-B induction. In search of additional signaling pathways activated by NIK, we now demonstrate that NIK activates MEK1 phosphorylation and induces the Erk1/Erk2 MAPK pathway. Treatment of PC12 cells with PD98059, a MEK1 inhibitor, potently blocked neurite process formation; however, a dominantly interfering mutant of the upstream Shc adapter failed to alter this response. These findings reveal a new function for NIK as a MEK1-dependent activator of the MAPK pathway and implicate both the IB kinase and MAPK signaling cascades in NIK-induced differentiation of PC12 cells.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

The NF-B-inducing kinase (NIK)¹ is a mitogen-activated protein kinase kinase kinase that potently induces NF-B. Although earlier studies placed NIK in the TNF/TNFR1 signaling cascade, more recent studies in NIK-deficient animals do not support this assignment (1, 2).² Although the upstream receptor remains uncertain, downstream NIK signaling leads to marked activation of the IB kinases (IKKs) (3), which phosphorylate IB, leading to its rapid ubiquitination and degradation in the 26 S proteasome. Inactive NF-B/B may continuously shuttle between the cytoplasm and nucleus; however, degradation of IB allows DNA binding by the liberated NF-B complex and promotes target gene expression (4-11). NF-B plays a key role in cytokine and growth factor signaling, serving to regulate the expression of a panoply of genes that mediate inflammatory, anti-apoptotic, and proliferation/differentiation signaling (12, 13).

The function of NF-B in the nervous system remains poorly understood. NF-B induction is associated with central nervous system lesions in such diverse diseases as Parkinson's (14), Alzheimer's (15), AIDS dementia (16), and spinal cord injury (17). This activation may be principally a protective response by the host to enhance resistance to neuronal insults and modulate synaptic plasticity (18). Mice lacking the NF-B family member p50 are more sensitive to kainic acid-induced excitotoxicity (19). Blocking NF-B activation increases the sensitivity of neuronal cells to apoptosis induced in response to reactive oxygen species and TNF (20). The involvement of NIK in the central nervous system is revealed by a naturally occurring autosomal recessive mutation in the alymphoplasia (aly) allele of mice. Aly mice contain a point mutation in the carboxyl terminus of the NIK gene that affects its ability to interact with TNF receptor-associated factors (2). These mice are immunodeficient because of a lack of lymph nodes but also develop progressive neurological abnormalities leading to hind limb paralysis (21). Interestingly, the gene encoding human NIK maps to chromosome 17 and more specifically to a locus associated with frontotemporal dementia and parkinsonism (22). Here we describe a series of studies examining intracellular signaling and responses in PC12 cells engineered to inducibly express NIK. PC12 cells are useful for the study of signaling events associated with neuronal survival, proliferation, and neuritogenesis (23, 24). We demonstrate that the ecdysone-induced expression of kinase-proficient but not -deficient NIK promotes neurite formation, involving activation of the IKK and MAPK pathways.

	EXPERIMENTAL PROCEDURES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

Expression Vectors and Cell Culture-- The expression vectors encoding IB S32A/S36A (SS/AA) and the dominant-negative Shc (Y239F/Y240F/Y317F) mutant have been described elsewhere (25, 26). Ecdysone-responsive (EcR) PC12 cells stably expressing WT Myc-NIK and kinase-deficient Myc-NIK (K429A/K430A) under the control of the ecdysone-inducible pIND vector (Invitrogen) were selected with hygromycin (Life Technologies, Inc.). Parental PC12 EcR cells and those stably transfected with the pIND Myc-NIK constructs were cultured in Dulbecco's modified Eagle's medium supplemented with 10% horse serum, 5% fetal bovine serum, and 1% penicillin/streptomycin (Life Technologies, Inc.) at 37 °C in a humidified atmosphere with 5% CO₂. Cells were treated as indicated with ecdysone (5 µM) (Invitrogen), murine TNF (20 ng/ml) (Sigma), mNGF (50 ng/ml) (Sigma), and the MEK1 inhibitor, PD98059 (50 µM) (New England Biolabs).

Immunoprecipitation and Immunoblotting Analyses-- EcR WT NIK PC12 cells, EcR kinase-deficient (KD) NIK PC12 cells, and parental EcR PC12 cells were stimulated with ecdysone (5 µM) for 24 h. The cells were washed with phosphate-buffered saline and lysed in eukaryotic lysis buffer (1% Nonidet P-40, 50 mM HEPES, pH 7.4, 250 mM NaCl, 5 mM EDTA, 10% glycerol, 0.5 mM DTT, 2 µg/ml aprotinin, 1 µg/ml leupeptin, and 1 mM phenylmethylsulfonyl fluoride). Cleared lysates were incubated with agarose-conjugated anti-myc rabbit polyclonal antibodies (Santa Cruz Biotechnology), and bound material was analyzed by SDS-PAGE, transferred to PVDF membranes, and immunoblotted with anti-myc antibodies.

Reporter Assay-- Approximately 2.5 × 10⁵ PC12 cells per well in six-well dishes were transfected with Lipofectin (Life Technologies, Inc.) and a luciferase reporter plasmid DNA containing B, AP1, CRE, or serum-response element (SRE) enhancer elements (Stratagene). After the indicated treatment and lysis in eukaryotic lysis buffer, luciferase activity was measured with the enhanced luciferase assay kit and a Microbeta 1450 Trilux luminescence counter (Wallac Company). All transfections included the pRC--actin LacZ plasmid to normalize for differences in gene transfer efficiency.

Apoptosis Assay and PC12 Cell Differentiation-- Cleavage of genomic DNA occurring during apoptosis was detected in TUNEL assays with terminal deoxynucleotidyl transferase, which catalyzes the polymerization of nucleotides to free 3' ends of DNA in a template-independent manner. Fluorescein labels incorporated in nucleotide polymers were detected and quantitated by fluorescence microscopy (in situ cell death detection kit; Roche Molecular Biochemicals). PC12 cell differentiation and survival was measured by plating cells onto collagen-coated eight-chamber glass culture slides (Falcon), and the percentage of cells with neurite processes greater than two cell bodies in length were counted after relevant treatment.

Kinase Assay-- EcR WT NIK PC12 cells, EcR KD NIK PC12 cells, and parental EcR PC12 cells were stimulated with ecdysone (5 µM) for 24 h and lysed in eukaryotic cell lysis buffer. Lysates were immunoprecipitated with rabbit polyclonal anti-IKK (H744, cross-reactive with IKK; Santa Cruz Biotechnology) and protein A-Sepharose (Amersham Pharmacia Biotech). The immune complexes were washed three times in lysis buffer and once in kinase buffer, resuspended in 20 µl of kinase buffer (20 mM HEPES, pH 7.4, 2 mM MnCl₂, 10 mM MgCl₂, 25 mM glycerol-2-phosphate, 0.1 mM Na₃VO₄, 4 mM NaF, 1 mM DTT, 20 µM ATP), and incubated with 5 µCi of [-³²P]ATP (6,000 Ci/mmol) and 1 µg of recombinant GST-IB (1-62) as an exogenous substrate. After incubation at 30 °C for 30 min, the kinase reactions were terminated by the addition of SDS-PAGE sample buffer. The samples were analyzed by SDS-PAGE, transferred to PVDF membrane, and exposed to hyperfilm (Amersham Pharmacia Biotech). The membranes were subsequently immunoblotted with anti-IKK to determine the relative amount of immunoprecipitated kinase. Activated MEK1 was measured by immunoblotting with an anti-phosphorylated MEK1/2 antibody (New England Biolabs). Expression of this kinase was measured by blotting with anti-MEK1 antibody (Santa Cruz Biotechnology).

Electrophoretic Mobility Shift Assay-- Nuclear extracts were prepared, and electrophoretic mobility shift assays were performed with a consensus B oligo (GGGGACTTTCCC) as a probe (Santa Cruz Biotechnology).

Atlas cDNA Array-- Atlas 1.2 cDNA arrays (CLONTECH) were used according to the manufacturer's protocol to assess the changes in gene expression in EcR WT NIK PC12 cells that were untreated or treated with ecdysone (5 µM) for 18 h. Probes are synthesized by reverse transcribing 1 µg of each RNA population using the cDNA synthesis primer mix and -³²P. Each radioactively labeled probe mix (1 × 10⁶ cpm) is then hybridized to separate Atlas arrays. After high stringency washes the hybridization pattern can be analyzed by autoradiography and quantitated by phosphorimaging.

	RESULTS AND DISCUSSION

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

To examine the effects of NIK, a potent upstream inducer of NF-B, on the survival and differentiation of PC12 cells, we prepared PC12 cells containing a stably integrated, EcR kinase-proficient (WT) or KD NIK expression plasmid. NIK expression was not detected under basal conditions; however, the addition of ecdysone (5 µM) to either the EcR WT NIK PC12 cells or the EcR KD NIK PC12 cells readily stimulated the expression of these gene products. As expected, the lysates from the parental EcR PC12 cells did not inducibly express NIK (Fig. 1A).

View larger version (49K): [in this window] [in a new window]   Fig. 1.   NIK expression and NF-B activity in PC12 cells. A, EcR WT NIK PC12 cells, EcR KD NIK PC12 cells, and parental EcR PC12 cells were stimulated with medium or ecdysone (Ecd) (5 µM) for 24 h at 37 °C. Myc-tagged NIK was immunoprecipitated with anti-Myc-conjugated agarose, and lysates were examined for the expression of NIK with anti-Myc antibodies. B, a B-luciferase reporter plasmid (B-luciferase) and a -galactosidase expression plasmid were cotransfected into EcR WT NIK PC12 cells, EcR KD NIK PC12 cells, and parental EcR PC12 cells. The cell lines were then treated with medium or Ecd (5 µM) for 24 h. B-luciferase activity was measured in the resultant lysates and normalized for -galactosidase activity to correct for differences in transfection efficiency between the cultures. -Fold induction for each sample was calculated relative to the value obtained with cells cultured with medium alone. Error bars indicate standard deviation derived from triplicate samples. C, EcR WT NIK PC12 cells, EcR KD NIK PC12 cells, and parental EcR PC12 cells were stimulated with Ecd (5 µM) for 24 h, and EcR NIK PC12 cells were treated with medium or Ecd (5 µM) for 24 h, and cellular lysates were prepared. IKK was immunoprecipitated from each lysate, suspended in kinase buffer, and subjected to an in vitro kinase assay employing GST-LB (1-62) as an exogenously added substrate. Samples were analyzed by SDS-PAGE for evidence of phosphorylation of GST-IB (1-62) and immunoblotted (IB) with anti-IKK antibodies to confirm the presence of comparable amounts of kinase in each reaction. D, EcR WT NIK PC12 cells were treated with medium alone or Ecd (5 µM) for 6, 18, and 24 h, and nuclear extracts were prepared. Electrophoretic mobility shift of a labeled consensus NF-B-binding oligonucleotide was measured.

To assess the ability of the inducibly expressed WT and KD NIK proteins to activate NF-B in PC12 cells, we transiently transfected a B-luciferase reporter plasmid together with a -galactosidase expression plasmid to normalize for differences in transfection efficiency. The EcR WT NIK, EcR KD NIK, and parental EcR PC12 cells were then stimulated with ecdysone (5 µM) for 24 h. Expression of WT NIK increased activity of the B-luciferase reporter approximately 20-fold. Ecdysone did not increase luciferase activity in EcR KD NIK or parental EcR PC12 cells (Fig. 1B).

Although some nucleo-cytoplasmic shuttling may occur, NF-B present in complexes with IB predominantly resides in the cytoplasm. After cellular stimulation and activation of the IKKs, IB is phosphorylated, ubiquitinated, and rapidly degraded by the 26 S proteasome, freeing NF-B for DNA binding in the nucleus (4-11, 27-30). We stimulated EcR WT NIK PC12 cells, EcR KD NIK PC12 cells, and parental EcR PC12 cells with ecdysome (5 µM for 24 h) and measured the activity of the endogenous IKK complex in in vitro kinase assays using GST-IB (1-62) as an exogenously added substrate. IB phosphorylation was substantially increased in WT NIK cells, but not in the KD NIK or parental PC12 cells (Fig. 1C). Consistent with these results, we demonstrated that nuclear extracts from EcR WT NIK PC12 cells stimulated with ecdysone for 6, 18, and 24 h display B-DNA binding activity at the latter two time points (Fig. 1D). This slow and sustained response presumably reflects the kinetics of NIK protein synthesis following ecdysone stimulation.

Growing evidence indicates that NF-B exerts anti-apoptotic effects in neuronal cells and that a blockade of NF-B activation is frequently associated with increased apoptosis (31-35). We assessed whether the induced expression of NIK could prevent apoptosis occurring in response to serum starvation. Apoptosis was measured using the TUNEL assay. EcR WT NIK and EcR KD NIK PC12 cell lines were cultured in the presence or absence of ecdysone (5 µM) for 24 h and then incubated for 24 h in medium containing 1% horse serum (Fig. 2). Significant numbers of TUNEL-positive cells were observed in KD NIK PC12 cultures in the presence or absence of ecdysone induction. Similarly in the absence of ecdysone stimulation, EcR WT NIK PC12 cells display marked TUNEL-positive staining. In contrast, the induced expression of WT NIK protected the PC12 cells from apoptosis accompanying serum starvation. Of note, WT NIK PC12 cells also formed neurite processes, a response not seen in unstimulated cells or cells expressing KD NIK (Fig. 2; compare panel H with panels D and G).

View larger version (72K): [in this window] [in a new window]   Fig. 2.   NIK promotes neurite outgrowth and mediates anti-apoptotic signaling. EcR WT NIK (panels B, D, F, and H) and EcR KD NIK (Panels A, C, E, and G) PC12 cells were incubated with (panels E-H) or without (panels A-D) Ecd (5 µM) for 24 h. The tissue culture medium was then replaced with medium containing 1% horse serum for an additional 24 h. Apoptotic cells were measured by TUNEL staining and fluorescence photomicroscopy (panels A, B, E, and F). Brightfield photomicroscopy allowed assessment of neurite formation in those cells also stained with TUNEL reagents (panels C, D, G, and H).

To explore whether neurite formation depended on NF-B, we transiently cotransfected EcR WT NIK PC12 cells with expression plasmids encoding a nondegradable IB mutant (IB SS/AA), green fluorescent protein (GFP) to mark transfected cells, and a B-luciferase reporter to assess the completeness of NF-B inhibition by the IB mutant (Fig. 3). IB SS/AA inhibited neurite formation by nearly 50% (left panel) under conditions that completely suppressed B-luciferase induction (right panel). These results suggest that NF-B plays a role in neurite formation but that other factors and pathways must be involved, as well.

View larger version (10K): [in this window] [in a new window]   Fig. 3.   NIK-mediated neuritogenesis is partially blocked by IB SS/AA. EcR WT NIK PC12 cells were transiently cotransfected with B-luciferase, -galactosidase, and pEGFP plasmids. Vector alone or 0.5 µg of an expression plasmid encoding the IB SS/AA mutant was also transfected. The effects of ectopically expressed IB (SS/AA) are presented as a percentage of the NIK-induced B-luciferase activity normalized for -galactosidase activity in ecdysone-treated cells transfected with only the reporter vector and the appropriate empty vector DNA. GFP-positive cells with neurites greater than two-cell bodies-long were scored as differentiated. Error bars indicate standard deviation derived from triplicate experiments.

Because NF-B induction may lead to NGF production (36, 37), we considered the possibility that NIK differentiates PC12 cells by an autocrine mechanism. However, transient expression of a dominant-negative Shc analogue (Shc YYY/FFF) failed to block WT NIK-mediated neurite formation even though it effectively inhibited NGF-stimulated differentiation (data not shown). Additionally, pretreatment of EcR WT NIK PC12 cells with anti-NGF antiserum had no effect on WT NIK-mediated neurite formation, and transfer of conditioned medium from NIK-expressing cells to new cultures failed to induce differentiation (data not shown). Together, these findings argue against an autocrine mechanism of NIK action involving the induced secretion of NGF.

Several transcription factors have been implicated in the differentiation of PC12 cells, including Fos, Jun, Elk, and CREB (38). Fos and Jun form homo- or hetero-dimers and bind to AP1 sites, Elk binds to SRE, and CREB is able to bind to CRE sites (39). To assess the involvement of these signaling pathways in WT NIK-mediated differentiation of PC12 cells, we tested the ability of WT NIK to activate AP1-, CRE-, or SRE-luciferase reporter plasmids. In addition to activating NF-B, ecdysone (5 µM for 24 h)-induced WT NIK expression also activated the SRE-luciferase reporter, suggesting involvement of the MEK1 pathway (Fig. 4A). Conversely WT NIK expression did not activate AP1-luciferase and stimulated minimal CRE-luciferase activity. As expected, stimulation of PC12 cells with NGF (50 ng/ml for 24 h) activated all three pathways, and TNF (20 ng/ml for 24 h) stimulation produced an AP1-luciferase response but failed to enhance CRE- or SRE-luciferase activity.

View larger version (47K): [in this window] [in a new window]   Fig. 4.   NIK activates the MEK1 signaling pathway. A, EcR WT NIK PC12 cells were cotransfected with AP1-, CRE-, or SRE-luciferase reporter plasmid in combination with a -galactosidase expression plasmid. The cells were then treated for 24 h with medium, TNF (20 ng/ml), NGF (50 ng/ml), or Ecd (5 µM), lysed, and examined for luciferase activity and normalized to -galactosidase activity. B, EcR WT NIK PC12 cells were transfected with a -galactosidase expression plasmid and the SRE-luciferase reporter plasmid. The effects of ectopically expressed dominant-negative Shc (Y317F/Y239F/Y240F) on NIK (5 µM Ecd/24 h)- or NGF (50 ng/ml/24 h)-mediated SRE-luciferase are shown. C, EcR WT NIK PC12 cells were incubated with media or Ecd (5 µM/24 h) or Ecd (5 µM/24 h) after a 1-h pretreatment with the MEK1 inhibitor PD98059 (50 µM). Cellular lysates were prepared and analyzed by SDS-PAGE and immunoblotted (IB) with anti-phosphorylated MEK1 to assess activity and anti-MEK1 to compare loading. D, EcR WT NIK PC12 cells were treated with medium or Ecd (5 µM) for 24 h or were pretreated with the MEK1 inhibitor PD98059 (50 µM) and then treated with Ecd (5 µM) for 24 h. The cells were then fixed and photographed.

To explore the pathway leading to SRE-luciferase activation, we tested the ability of a dominantly interfering Shc mutant (YYY/FFF) to block NIK- and NGF-induced SRE-luciferase activity. This Shc mutant failed to inhibit the NIK response but significantly attenuated the NGF response (Fig. 4B). Of note, pretreatment of WT NIK PC12 cells with the MEK1 inhibitor PD98059 markedly inhibited both WT NIK- and NGF-mediated activation of SRE-luciferase activity.

To further test the involvement of MEK1 in the WT NIK response, we used a phospho-specific MEK1 antibody to monitor activation of this kinase (Fig. 4C). Induction of WT NIK expression led to phosphorylation of MEK1, and this response was blocked by the MEK1 inhibitor PD98059 (5 µM for 1 h). Probing the blot with anti-MEK1 confirmed equal loading of the lysates.

MEK1 activity has been shown to be necessary and sufficient for PC12 cell differentiation (40, 41). Therefore we tested whether MEK1 activation is required for WT NIK-mediated PC12 cell differentiation; we pretreated EcR WT NIK PC12 cells with PD98059 (50 µM) for 1 h before stimulation with ecdysone for 24 h. PD98059 effectively blocked differentiation of PC12 cells expressing WT NIK (Fig. 4D). These observations suggest that NIK acts downstream of Shc and upstream of MEK1. Because the activation loop sequences found in IKK, IKK, and MEK1 are quite similar in structure, each might serve as a substrate for NIK (42). These data indicate that NIK exerts a broader range of effects than previously suspected, activating both the IKK complex and MEK1.

We next surveyed WT NIK-mediated gene expression in PC12 cells using an Atlas cDNA array. Labeled cDNA probes from unstimulated EcR WT NIK PC12 cells and those treated with ecdysone for 18 h were hybridized to an Atlas 1.2 rat cDNA array, containing 588 different cDNAs. The genes up-regulated in response to NIK expression in PC12 cells included IB, cyclin D3, GSTµ, and the neuronal acetylcholine receptor protein (Table I). These data provide potential insights into the underlying mechanism by which NIK modulates signal regulation, cell-cycle progression, survival pathways, and neuronal cell function.

Together, our findings raise the possibility that defective NIK function in Aly mice contributes to the progressive neurological degeneration observed in these animals. We hypothesize that cells from these mice do not respond normally when exposed to a ligand capable of activating NIK. In the absence of NF-B to prevent neuronal apoptosis and the neuronal differentiation signals generated via both IKK- and MEK1-dependent pathways, these mice may develop progressive neurological deficits. A complete analysis of the neurological defect in the aly animals is currently underway.

	ACKNOWLEDGEMENTS

We thank John C. W. Carroll, Neile Shea, Chris Goodfellow, and Stephen Gonzales for assistance with graphics and Stephen Ordway and Gary Howard for excellent editorial input. We thank Eugene M. Johnson for providing anti-NGF antibody.

	FOOTNOTES

^* This work was supported in part by University of California at San Francisco Gladstone Institute of Virology and Immunology, Center for AIDS Research grant P30-MH59037.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^§ To whom correspondence should be addressed. Tel.: 415-695-3838; Fax: 415-826-1514; E-mail: efoehr@gladstone.ucsf.edu.

Published, JBC Papers in Press, September 8, 2000, DOI 10.1074/jbc.C000507200

² D. Goeddell, personal communication.

	ABBREVIATIONS

The abbreviations used are: NIK, NF-B-inducing kinase; TNF, tumor necrosis factor; IKK(s), IB kinase; MAPK, mitogen-activated protein kinase; EcR, ecdysone-responsive; WT, wild type; MEK, mitogen-activated protein kinase/extracellular signal-regulated kinase kinase; KD, kinase-deficient; DTT, dithiothreitol; PAGE, polyacrylamide gel electrophoresis; PVDF, polyvinylidene difluoride; CRE, cAMP-response element; SRE, serum-response element; TUNEL, terminal dUTP nick-end labeling; GST, glutathione S-transferase; GFP, green fluorescent protein; NGF, nerve growth factor; CREB, cAMP-response element-binding protein; Ecd, ecdysone.

	REFERENCES

TOP ABSTRACT INTRODUCTION EXPERIMENTAL PROCEDURES RESULTS AND DISCUSSION REFERENCES

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				W.-H. Hu, J. S. Pendergast, X.-M. Mo, R. Brambilla, V. Bracchi-Ricard, F. Li, W. M. Walters, B. Blits, L. He, S. M. Schaal, et al. NIBP, a Novel NIK and IKK{beta}-binding Protein That Enhances NF-{kappa}B Activation J. Biol. Chem., August 12, 2005; 280(32): 29233 - 29241. [Abstract] [Full Text] [PDF]

				A. Denys, A. Hichami, and N. A. Khan n-3 PUFAs modulate T-cell activation via protein kinase C-{alpha} and -{varepsilon} and the NF-{kappa}B signaling pathway J. Lipid Res., April 1, 2005; 46(4): 752 - 758. [Abstract] [Full Text] [PDF]

				S. A. Williams{section}, L.-F. Chen, H. Kwon, D. Fenard, D. Bisgrove, E. Verdin, and W. C. Greene Prostratin Antagonizes HIV Latency by Activating NF-{kappa}B J. Biol. Chem., October 1, 2004; 279(40): 42008 - 42017. [Abstract] [Full Text] [PDF]

				H. Rangaswami, A. Bulbule, and G. C. Kundu Nuclear Factor-inducing Kinase Plays a Crucial Role in Osteopontin-induced MAPK/I{kappa}B{alpha} Kinase-dependent Nuclear Factor {kappa}B-mediated Promatrix Metalloproteinase-9 Activation J. Biol. Chem., September 10, 2004; 279(37): 38921 - 38935. [Abstract] [Full Text] [PDF]

				W.-H. Hu, X.-M. Mo, W. M. Walters, R. Brambilla, and J. R. Bethea TNAP, a Novel Repressor of NF-{kappa}B-inducing Kinase, Suppresses NF-{kappa}B Activation J. Biol. Chem., August 20, 2004; 279(34): 35975 - 35983. [Abstract] [Full Text] [PDF]

				Y. Wu, W. Sheng, L. Chen, H. Dong, V. Lee, F. Lu, C. S. Wong, W.-Y. Lu, and B. B. Yang Versican V1 Isoform Induces Neuronal Differentiation and Promotes Neurite Outgrowth Mol. Biol. Cell, May 1, 2004; 15(5): 2093 - 2104. [Abstract] [Full Text] [PDF]

				K.-S. Chun, Y.-S. Keum, S. S. Han, Y.-S. Song, S.-H. Kim, and Y.-J. Surh Curcumin inhibits phorbol ester-induced expression of cyclooxygenase-2 in mouse skin through suppression of extracellular signal-regulated kinase activity and NF-{kappa}B activation Carcinogenesis, September 1, 2003; 24(9): 1515 - 1524. [Abstract] [Full Text] [PDF]

				L. E. Jensen and A. S. Whitehead Pellino3, a Novel Member of the Pellino Protein Family, Promotes Activation of c-Jun and Elk-1 and May Act as a Scaffolding Protein J. Immunol., August 1, 2003; 171(3): 1500 - 1506. [Abstract] [Full Text] [PDF]

				J. F. Sanchez, L. F. Sniderhan, A. L. Williamson, S. Fan, S. Chakraborty-Sett, and S. B. Maggirwar Glycogen Synthase Kinase 3{beta}-Mediated Apoptosis of Primary Cortical Astrocytes Involves Inhibition of Nuclear Factor {kappa}B Signaling Mol. Cell. Biol., July 1, 2003; 23(13): 4649 - 4662. [Abstract] [Full Text] [PDF]

				C. L. Crowley-Weber, C. M. Payne, M. Gleason-Guzman, G. S. Watts, B. Futscher, C. N. Waltmire, C. Crowley, K. Dvorakova, C. Bernstein, M. Craven, et al. Development and molecular characterization of HCT-116 cell lines resistant to the tumor promoter and multiple stress-inducer, deoxycholate Carcinogenesis, December 1, 2002; 23(12): 2063 - 2080. [Abstract] [Full Text] [PDF]

				P. Dhawan and A. Richmond A Novel NF-kappa B-inducing Kinase-MAPK Signaling Pathway Up-regulates NF-kappa B Activity in Melanoma Cells J. Biol. Chem., March 1, 2002; 277(10): 7920 - 7928. [Abstract] [Full Text] [PDF]

				M. A. Luftig, E. Cahir-McFarland, G. Mosialos, and E. Kieff Effects of the NIK aly Mutation on NF-kappa B Activation by the Epstein-Barr Virus Latent Infection Membrane Protein, Lymphotoxin beta Receptor, and CD40 J. Biol. Chem., April 27, 2001; 276(18): 14602 - 14606. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) All Versions of this Article: 275/44/34021    most recent C000507200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Foehr, E. D. Articles by Greene, W. C. Search for Related Content PubMed PubMed Citation Articles by Foehr, E. D. Articles by Greene, W. C. Social Bookmarking                      What's this?