Eriodictyol Inhibits RSK2-ATF1 Signaling and Suppresses EGF-induced Neoplastic Cell Transformation

Hormel Institute, University of Minnesota, Austin, Minnesota 55912, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 01/2011; 286(3):2057-66. DOI: 10.1074/jbc.M110.147306
Source: PubMed


RSK2 is a widely expressed serine/threonine kinase, and its activation enhances cell proliferation. Here, we report that ATF1 is a novel substrate of RSK2 and that RSK2-ATF1 signaling plays an important role in EGF-induced neoplastic cell transformation. RSK2 phosphorylated ATF1 at Ser-63 and enhanced ATF1 transcriptional activity. Docking experiments using the crystal structure of the RSK2 N-terminal kinase domain combined with in vitro pulldown assays demonstrated that eriodictyol, a flavanone found in fruits, bound with the N-terminal kinase domain of RSK2 to inhibit RSK2 N-terminal kinase activity. In cells, eriodictyol inhibited phosphorylation of ATF1 but had no effect on the phosphorylation of RSK, MEK1/2, ERK1/2, p38 or JNKs, indicating that eriodictyol specifically suppresses RSK2 signaling. Furthermore, eriodictyol inhibited RSK2-mediated ATF1 transactivation and tumor promoter-induced transformation of JB6 Cl41 cells. Eriodictyol or knockdown of RSK2 or ATF1 also suppressed Ras-mediated focus formation. Overall, these results indicate that RSK2-ATF1 signaling plays an important role in neoplastic cell transformation and that eriodictyol is a novel natural compound for suppressing RSK2 kinase activity.

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    • "However, a recent study found that in mice with targeted deletion of all three members of the miR-34 family, the p53 response was not impaired in a variety of in vivo and in vitro assays, indicated that miR-34 members are not critical for downstream effectors of p53 [49]. Inhibition of miR-34c could prevent mouse male germ cell apoptosis through targeting ATF1 (activating transcription factor 1) [50], which mediates the transcriptional response of various extracellular signals and it is involved in cell viability and cell transformation [51]–[54], providing a novel mechanism with involvement of miRNAs in the regulation of germ cell apoptosis. "
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    ABSTRACT: MicroRNAs (miRNAs) are the class of small endogenous RNAs that play an important regulatory role in cells by negatively affecting gene expression at transcriptional and post-transcriptional levels. There have been extensive studies aiming to discover miRNAs and to analyze their functions in the cells from a variety of species. However, there are no published studies of miRNA profiles in human testis using next generation sequencing (NGS) technology. We employed Solexa sequencing technology to profile miRNAs in normal human testis. Total 770 known and 5 novel human miRNAs, and 20121 piRNAs were detected, indicating that the human testis has a complex population of small RNAs. The expression of 15 known and 5 novel detected miRNAs was validated by qRT-PCR. We have also predicted the potential target genes of the abundant known and novel miRNAs, and subjected them to GO and pathway analysis, revealing the involvement of miRNAs in many important biological phenomenon including meiosis and p53-related pathways that are implicated in the regulation of spermatogenesis. This study reports the first genome-wide miRNA profiles in human testis using a NGS approach. The presence of large number of miRNAs and the nature of their target genes suggested that miRNAs play important roles in spermatogenesis. Here we provide a useful resource for further elucidation of the regulatory role of miRNAs and piRNAs in the spermatogenesis. It may also facilitate the development of prophylactic strategies for male infertility.
    PLoS ONE 06/2013; 8(6):e66809. DOI:10.1371/journal.pone.0066809 · 3.23 Impact Factor
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    • "In an effort to identify new RSK2 specific inhibitors, Liu et al (2011) identified eriodictyol through molecular docking [13]. Interestingly, eriodictyol is a flavonoid that is structurally very similar to, luteolin, apigenin and kaempferol. "
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    Oncotarget 02/2013; 4(2):329-45. DOI:10.18632/oncotarget.834 · 6.36 Impact Factor
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    • "Activating transcription factor 1 (ATF1), which constitutes a subfamily of the basic leucine zipper transcription factors, mediates the transcriptional response of various extracellular signals and it is involved in cell viability and cell transformation [22], [23], [24], [25]. In human clear cell sarcoma, the ATF1 gene seems to be responsible for maintaining tumor viability [26]. "
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    ABSTRACT: MicroRNAs (miRNAs) play vital regulatory roles in many cellular processes. The expression of miRNA (miR)-34c is highly enriched in adult mouse testis, but its roles and underlying mechanisms of action are not well understood. In the present study, we show that miR-34c is detected in mouse pachytene spermatocytes and continues to be highly expressed in spermatids. To explore the specific functions of miR-34c, we have established an in vivo model by transfecting miR-34c inhibitors into primary spermatocytes to study the loss-of-function of miR-34c. The results show that silencing of miR-34c significantly increases the Bcl-2/Bax ratio and prevents germ cell from apoptosis induced by deprivation of testosterone. Moreover, ectopic expression of the miR-34c in GC-2 cell trigger the cell apoptosis with a decreased Bcl-2/Bax ratio and miR-34c inhibition lead to a low spontaneous apoptotic ratio and an increased Bcl-2/Bax ratio. Furthermore, ectopic expression of miR-34c reduces ATF1 protein expression without affecting ATF1 mRNA level via directly binding to ATF1's 3'UTR, indicating that ATF1 is one of miR-34c's target genes. Meanwhile, the knockdown of ATF1 significantly decreases the Bcl-2/Bax ratio and triggers GC-2 cell apoptosis. Inhibition of miR-34c does not decrease the GC-2 cell apoptosis ratio in ATF1 knockdown cells. Our study shows for the first time that miR-34c functions, at least partially, by targeting the ATF1 gene in germ cell apoptosis, providing a novel mechanism with involvement of miRNA in the regulation of germ cell apoptosis.
    PLoS ONE 03/2012; 7(3):e33861. DOI:10.1371/journal.pone.0033861 · 3.23 Impact Factor
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