Biochemical characterization of mouse brain necdin

Maruyama, Etsuko

doi:10.1042/bj3140895

Cited by 17 publications

(16 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Necdin is a cell-growth suppressor, and has function similar to that of tumor suppressor Rb (37). Necdin, itself may be a DNA-binding protein, because it shows tight binding on dsDNA affinity chromatography (38). Our results suggest that the binding site of necdin with IL-1␣ resides within the N-terminal region of necdin.…”

Section: Discussionmentioning

confidence: 62%

A nuclear target for interleukin-1α: Interaction with the growth suppressor necdin modulates proliferation and collagen expression

Wang

Zhang

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

There is growing evidence for the intracellular role of cytokines and growth factors, but the pathways by which these activities occur remain largely obscure. Previous work from our laboratory identified the constitutive, aberrant expression of the 31-kDa IL-1␣ precursor (pre-IL-1␣) in the nuclei of fibroblasts from the lesional skin of patients with systemic sclerosis (SSc). We established that pre-IL-1␣ expression was associated with increased fibroblast proliferation and collagen production. Further investigation has led to the identification of a mechanism by which nuclear expression of pre-IL-1␣ affects fibroblast growth and matrix production. By using a yeast two-hybrid method, we found that pre-IL-1␣ binds necdin, a nuclear protein with growth suppressor activity. We mapped the region of pre-IL-1␣ responsible for necdin binding and found it to be localized near the N terminus, a region that is present on pre-IL-1␣, but not the mature 17-kDa cytokine. Expression studies demonstrated that pre-IL-1␣ associates with necdin in the nuclei of mammalian cell lines and regulates cell growth and collagen expression. Our results provide the first evidence, to our knowledge, of a nuclear target for pre-IL-1␣. Based on these findings, we propose that the constitutively up-regulated expression of pre-IL-1␣ in the nuclei of SSc fibroblasts up-regulates proliferation and matrix production of SSc fibroblasts through binding necdin, and by counteracting its effects on cell growth and collagen production. IL-1␣ is a pleiotropic cytokine that has numerous effects on the growth and activation on a wide variety of cell types (1). Pre-IL-1␣ is the precursor of mature IL-1␣ containing a nuclear localization sequence (NLS) in its N-terminal region. Processing of the 31-kDa pre-IL-1␣ to the 17-kDa mature form requires the membrane-associated cysteine proteases called calpains, and a variety of cells appear to be deficient in this processing (1). Several studies (2-8) indicate that pre-IL-1␣ accumulates as an intracellular precursor molecule with a molecular mass of 31-kDa in epithelial cells (keratinocytes) and some mesenchymal cells (i.e., endothelial cells and fibroblasts). Previous reports indicate that intracellular pre-IL-1␣ may serve as a regulator of cell growth, senescence, and differentiation (2-9) through an intracrine mechanism (10-16), through interaction with as yet unidentified intracellular target(s).Fibrosis is one of the key features of systemic sclerosis (SSc), and is caused by increased proliferation and extracellular matrix production by fibroblasts of SSc patients (17,18). In contrast to healthy adult human skin fibroblasts, SSc fibroblasts are resistant to the induction of quiescence by serum deprivation, continuing to synthesize DNA, and to express the immediate response protooncogene, c-myc (19). Previous reports (17)(18)(20)(21)(22)(23)(24) have indicated that IL-1␣ increases fibroblast proliferation and collagen production. SSc fibroblasts constitutively express pre-IL-1␣ (2, 25), and the distribution of ...

show abstract

Section: Discussionmentioning

confidence: 62%

A nuclear target for interleukin-1α: Interaction with the growth suppressor necdin modulates proliferation and collagen expression

Wang

Zhang

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…13 For gluteraldehyde cross-linking studies, lysates were treated with 1 mM gluteraldehyde. 45 FLAG-or V5-CDC5L and mutants were immunoprecipitated as detailed above, and proteins were suspended in 0.1% bromophenol blue, 10% glycerol for electrophoretic separation in 4-20% polyacrylamide-TrisHCl gels under native conditions, or in 50 mM Tris-HCl, pH 6.8, 2% SDS, 0.1% bromophenol blue, 10% glycerol, 100 mM dithiothreitol for electrophoresis in 8% polyacrylamide-SDS gels.…”

Section: Homodimerization Assaymentioning

confidence: 99%

Cell cycle-dependent phosphorylation of human CDC5 regulates RNA processing

Gräub

Lancero²,

Pedersen³

et al. 2008

Cell Cycle

View full text Add to dashboard Cite

CDC5 proteins are components of the pre-mRNA splicing complex and essential for cell cycle progression in yeast, plants and mammals. Human CDC5 is phosphorylated in a mitogen-dependent manner, and its association with the spliceosome is ATP-dependent. Examination of the amino acid sequence suggests that CDC5L may be phosphorylated at up to 28 potential consensus recognition sequences for known kinases, however, the identity of actual phosphorylation sites, their role in regulating CDC5L activity, and the kinases responsible for their phosphorylation have not previously been determined. Using two-dimensional phosphopeptide mapping and nanoelectrospray mass spectrometry, we now show that CDC5L is phosphorylated on at least nine sites in vivo. We demonstrate that while CDC5L is capable of forming homodimers in vitro and in vivo, neither homodimerization nor nuclear localization is dependent on phosphorylation at these sites. Using an in vitro splicing assay, we show that phosphorylation of CDC5L at threonines 411 and 438 within recognition sequences for CDKs are required for CDC5L-mediated pre-mRNA splicing. We also demonstrate that a specific inhibitor of CDK2, CVT-313, inhibits CDC5L phosphorylation in both in vitro kinase assays and in vivo radiolabeling experiments in cycling cells. These studies represent the first demonstration of a regulatory role for phosphorylation of CDC5L, and suggest that targeting these sites or the implicated kinases may provide novel strategies for treating disorders of unguarded cellular proliferation, such as cancer.

show abstract

“…Indeed, Necdin was discovered because its expression was markedly increased when cancer cells were induced to differentiate (41). Necdin is recognized as a basic nuclear DNA binding protein that can associate with other transcription factors to regulate the activity of a number of cellular genes (40). However, the Necdin gene is more commonly associated with the neurogenetic disorder PWS, in which this gene is inactivated or deleted (36).…”

Section: Discussionmentioning

confidence: 99%

“…The localization of Necdin in differentiated neurons was previously demonstrated to show predominantly cytoplasmic signals, with a striking change in location to the nucleus during specific physiological changes (55). Necdin has also been shown to interact with DNA, which may enable it to regulate transcription of other genes involved in cellular differentiation and proliferation through direct binding with DNA or through its interaction with other transcription factors, including p53 and E2F1 (5,40). In fact, Necdin has been shown to act as a cellular transcriptional repressor by directly binding to multiple guanosine clusters within the promoters of target genes (43).…”

mentioning

confidence: 99%

EBNA3C Can Modulate the Activities of the Transcription Factor Necdin in Association with Metastasis Suppressor Protein Nm23-H1

Kaul

Murakami

Lan

et al. 2009

J Virol

View full text Add to dashboard Cite

Previous studies have demonstrated the interaction between the Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA3C) and the metastatic suppressorEpstein-Barr virus (EBV) is a human gammaherpesvirus which predominantly targets B cells and epithelial cells and is associated with a number of human cancers, including Burkitt's lymphoma, nasopharyngeal carcinoma, Hodgkin's disease, AIDS-associated and transplant-associated immunoblastic lymphoma, and, controversially, invasive breast carcinoma (9, 61). In vitro infection of B cells with EBV gives rise to lymphoblastoid cell lines (LCLs) which express a subset of 12 latent viral transcripts (61). These 12 transcripts encode six nuclear antigens (EBNAs), three latent membrane proteins, two early RNAs (32), and the BARF transcripts (61). Studies have shown that EBNA3C, one of the six nuclear antigens, interacts with the suppressor of cell migration and metastasis Nm23-H1 (69). This interaction between Nm23-H1 and EBNA3C has been shown to result in an increase in transcriptional activity on a targeted responsive promoter (70). Nm23-H1 tethered to DNA by a Gal4 DNA binding domain can activate transcription from a basal promoter at relatively low levels. However, when EBNA3C was introduced, the transactivation activity was shown to be substantially increased (70). These results suggest that Nm23-H1 may possess transcriptional regulatory activities which could function independent of its direct binding with DNA. This could possibly be through its interaction with EBNA3C (69). Interestingly, the presence of EBNA3C mediates the cellular translocation of Nm23-H1 from a mostly cytoplasmic to a predominantly nuclear signal. Moreover, EBNA3C can reverse the antimigratory effects of Nm23-H1 in vitro (69) as well as in vivo (23). The interaction between EBNA3C and Nm23-H1 has also been shown to be important in the regulation of COX-2, MMP-9, and alpha V integrin (13,24,26).The Nm23 gene family is highly conserved among a wide variety of eukaryotic species. Eight Nm23 genes have been identified for humans: Nm23-H1 (62), Nm23-H2 (68), Nm23-H3 (79), Nm23-H4 (45), Nm23-H5 (47), 76),. Members of the Nm23 gene family are structurally and functionally conserved, consisting of four to six identically folded subunits enclosing a large (25-Å) central cavity (84). Expression of Nm23 genes has been linked to suppression of tumor metastasis, differentiation, apoptosis, proliferation, and DNA mutation rate (14) and is also associated with nucleoside diphosphate (NDP) kinase activity (7,49,82), serine phosphorylation (8,20,37,48), histidine protein kinase activities (81), and transcriptional stimulatory activities (2,6,59,60). The human Nm23-H1 gene product is the best-characterized member of this family of proteins. It is 152 amino acids (aa) in length with leucine repeats and alpha-helical and basic domains (61). Nm23-H1 is 88% identical to Nm23-H2 and maps 4 kb apart at position q21.3 on

show abstract

Biochemical characterization of mouse brain necdin

Cited by 17 publications

References 12 publications

A nuclear target for interleukin-1α: Interaction with the growth suppressor necdin modulates proliferation and collagen expression

A nuclear target for interleukin-1α: Interaction with the growth suppressor necdin modulates proliferation and collagen expression

Cell cycle-dependent phosphorylation of human CDC5 regulates RNA processing

EBNA3C Can Modulate the Activities of the Transcription Factor Necdin in Association with Metastasis Suppressor Protein Nm23-H1

Contact Info

Product

Resources

About