title: Post-transcriptional regulation of BCL2 mRNA by the RNA-binding protein ZFP36L1 in malignant B cells.
abstract: The human ZFP36 zinc finger protein family consists of ZFP36, ZFP36L1, and ZFP36L2. These proteins regulate various cellular processes, including cell apoptosis, by binding to adenine uridine rich elements in the 3' untranslated regions of sets of target mRNAs to promote their degradation. The pro-apoptotic and other functions of ZFP36 family members have been implicated in the pathogenesis of lymphoid malignancies. To identify candidate mRNAs that are targeted in the pro-apoptotic response by ZFP36L1, we reverse-engineered a gene regulatory network for all three ZFP36 family members using the 'maximum information coefficient' (MIC) for target gene inference on a large microarray gene expression dataset representing cells of diverse histological origin. Of the three inferred ZFP36L1 mRNA targets that were identified, we focussed on experimental validation of mRNA for the pro-survival protein, BCL2, as a target for ZFP36L1. RNA electrophoretic mobility shift assay experiments revealed that ZFP36L1 interacted with the BCL2 adenine uridine rich element. In murine BCL1 leukemia cells stably transduced with a ZFP36L1 ShRNA lentiviral construct, BCL2 mRNA degradation was significantly delayed compared to control lentiviral expressing cells and ZFP36L1 knockdown in different cell types (BCL1, ACHN, Ramos), resulted in increased levels of BCL2 mRNA levels compared to control cells. 3' untranslated region luciferase reporter assays in HEK293T cells showed that wild type but not zinc finger mutant ZFP36L1 protein was able to downregulate a BCL2 construct containing the BCL2 adenine uridine rich element and removal of the adenine uridine rich core from the BCL2 3' untranslated region in the reporter construct significantly reduced the ability of ZFP36L1 to mediate this effect. Taken together, our data are consistent with ZFP36L1 interacting with and mediating degradation of BCL2 mRNA as an important target through which ZFP36L1 mediates its pro-apoptotic effects in malignant B-cells.
authors.affiliation: Division of Immunology, Infection and Inflammatory Disease, King's College London, London, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Zekavati
authors.affiliation: Division of Immunology, Infection and Inflammatory Disease, King's College London, London, United Kingdom.
authors.forename: Asghar
authors.initials: A
authors.lastname: Nasir
authors.affiliation: Department of Biomedical Sciences, University of Westminster, London, United Kingdom.
authors.forename: Amor
authors.initials: A
authors.lastname: Alcaraz
authors.affiliation: Division of Immunology, Infection and Inflammatory Disease, King's College London, London, United Kingdom.
authors.forename: Maceler
authors.initials: M
authors.lastname: Aldrovandi
authors.affiliation: Division of Endocrinology, King's College London, London, United Kingdom.
authors.forename: Phil
authors.initials: P
authors.lastname: Marsh
authors.affiliation: School of Biological Sciences, University of Essex, Colchester, Essex, United Kingdom.
authors.forename: John D
authors.initials: JD
authors.lastname: Norton
authors.affiliation: Division of Immunology, Infection and Inflammatory Disease, King's College London, London, United Kingdom; Department of Biomedical Sciences, University of Westminster, London, United Kingdom.
authors.forename: John J
authors.initials: JJ
authors.lastname: Murphy
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001402:B-Lymphocytes; D051817:Butyrate Response Factor 1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D011485:Protein Binding; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D020871:RNA Stability; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D015398:Signal Transduction
chemical_list: D020413:3' Untranslated Regions; C000595707:BCL2 protein, human; D051817:Butyrate Response Factor 1; D009687:Nuclear Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; C102034:ZFP36L1 protein, human; C488431:Zfp36l1 protein, mouse; C052617:Bcl2 protein, mouse; D008156:Luciferases