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Post‐transcriptional Control of Human Interleukin‐3 Mediated by its 3′‐UTR
Author(s) -
HernándezPérez Marimar,
Almodov Natalia,
GonzalezFeliciano Jose A,
Gonzalez Carlos
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.494.3
Subject(s) - luciferase , translational regulation , transfection , three prime untranslated region , untranslated region , hela , microbiology and biotechnology , messenger rna , biology , chemistry , translation (biology) , five prime untranslated region , cell culture , biochemistry , gene , genetics
Human interleukin‐3 (IL‐3) is a cytokine that stimulates the growth and differentiation of early lymphoid stem cells and has been implicated in certain types of cancer. IL‐3 is a member of a class of transiently expressed mRNAs that harbor Adenosine/Uridine‐Rich Elements (ARE) in their 3′‐UTRs. The 3′‐UTRs of many cytokines play a role in post‐transcriptional control by altering mRNA stability and/or translation. Transfection assays using HeLa and Jurkat cells harboring a luciferase‐IL‐3 3′‐UTR reporter showed a significant reduction in luciferase activity. The mRNA levels of these luciferase chimera do not show any significant differences in HeLa and T‐cells, suggesting that the hIL‐3 3′‐UTR mediates post‐transcriptional control. To understand how the AREs from the hIL‐3 3′‐UTR are involved in controlling translation, we conducted site‐directed mutagenesis of four ARE clusters that were identified in the hIL‐3 3′‐UTR. Firefly luciferase reporters harboring ARE mutations were transfected into HeLa and T cells. The luciferase chimeras lacking the hIL‐3 ARE showed an increase in luciferase activity. Electrophoretic mobility shift assays (EMSA) demonstrated that TIA‐1 and HuR, two ARE‐BP previously associated with translational control, bind to the hIL‐3 ARE. These studies will help delineate the functionality of AREs in the translational control of human IL‐3 and its relationship with oncogenesis. Supported by grants from NIH to C.I.G. (KO1 HL‐04355‐05, U54 CA96297, P20 RR 016174). M.H. is supported by the RISE Program, R25GM061838.