title: Lymphotoxin α, a novel target of posttranscriptional gene regulation by HuR in HepG2 cells.
abstract: The role of the RNA-binding protein human antigen R (HuR) in hepatocarcinogenesis is still elusive. By employing short hairpin (sh)RNA-dependent knockdown approach, we demonstrate that lymphotoxin α (LTα) is a target of posttranscriptional gene regulation by HuR in hepatocellular carcinoma (HepG2) cells. Consequently, the increased mRNA decay upon HuR depletion significantly affects lymphotoxin expression at both, the mRNA and protein level. Biotin-pulldown assay showed that HuR specifically interacts with the 3'-untranslated region (3'-UTR) of the LTα mRNA. Furthermore, electrophoretic mobility shift assay (EMSA) implicates that the RNA-binding critically depends on the RNA recognition motif 2 (RRM2) and the hinge region of HuR.
authors.affiliation: pharmazentrum frankfurt/ZAFES, Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany.
authors.forename: Nico
authors.initials: N
authors.lastname: Steinmeyer
authors.affiliation: Industrielle Biotechnologie, Fakultät Ingenieurwissenschaften, Hochschule Ansbach, Germany.
authors.forename: Anke
authors.initials: A
authors.lastname: Doller
authors.affiliation: pharmazentrum frankfurt/ZAFES, Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany.
authors.forename: Abhiruchi
authors.initials: A
authors.lastname: Biyanee
authors.affiliation: Institut für Biochemie I (Pathobiochemie), Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany.
authors.forename: Thilo
authors.initials: T
authors.lastname: Brauss
authors.affiliation: Institut für Biochemie I (Pathobiochemie), Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Schmid
authors.affiliation: pharmazentrum frankfurt/ZAFES, Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany.
authors.forename: Josef
authors.initials: J
authors.lastname: Pfeilschifter
authors.affiliation: pharmazentrum frankfurt/ZAFES, Klinikum der Goethe-Universität Frankfurt, Frankfurt am Main, Germany. Electronic address: w.eberhardt@em.uni-frankfurt.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eberhardt
mesh_terms: D020413:3' Untranslated Regions; D051959:ELAV Proteins / Q000235:genetics / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D056945:Hep G2 Cells; D006801:Humans; D008233:Lymphotoxin-alpha / Q000235:genetics* / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D020413:3' Untranslated Regions; D051959:ELAV Proteins; D008233:Lymphotoxin-alpha; D012333:RNA, Messenger
keywords: AU-rich element; Hepatocellular carcinoma; Lymphotoxin α; mRNA-stability