Genome-wide polysome profiling reveals an inflammation-responsive posttranscriptional operon in gamma interferon-activated monocytes.
We previously showed that ribosomal protein L13a is required for translational silencing of gamma interferon (IFN-gamma)-induced ceruloplasmin (Cp) synthesis in monocytes. This silencing also requires the presence of the GAIT (IFN-gamma activated inhibitor of translation) element in the 3' untranslated region (UTR) of Cp mRNA. Considering that Cp is an inflammatory protein, we hypothesized that this mechanism may have evolved to silence a family of proinflammatory proteins, of which Cp is just one member. To identify the other mRNAs that are targets for this silencing, we performed a genome-wide analysis of the polysome-profiled mRNAs by using an Affymetrix GeneChip and an inflammation-responsive gene array. A cluster of mRNAs encoding different chemokines and their receptors was identified as common hits in the two approaches and validated by real-time PCR. In silico predicted GAIT hairpins in the 3' UTRs of the target mRNAs were confirmed as functional cis-acting elements for translational silencing by luciferase reporter assays. Consistent with Cp, the newly identified target mRNAs also required L13a for silencing. Our studies have identified a new inflammation-responsive posttranscriptional operon that can be regulated directly at the level of translation in IFN-gamma-activated monocytes. This regulation of a cohort of mRNAs encoding inflammatory proteins may be important to resolve inflammation.
Vyas
Keyur
K
Center for Gene Regulation in Health and Disease and Department of Biology, Cleveland State University, Cleveland, Ohio 44115, USA.
Chaudhuri
Sujan
S
Leaman
Douglas W
DW
Komar
Anton A
AA
Musiyenko
Alla
A
Barik
Sailen
S
Mazumder
Barsanjit
B
D020413:3' Untranslated Regions; D002570:Ceruloplasmin / Q000378:metabolism; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D003198:Computer Simulation; D024202:Electrophoretic Mobility Shift Assay; D020868:Gene Silencing; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000378:metabolism; D007371:Interferon-gamma / Q000378:metabolism*; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D009876:Operon; D011132:Polyribosomes / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019707:Receptors, Chemokine / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D020298:U937 Cells
D020413:3' Untranslated Regions; D018925:Chemokines; D012333:RNA, Messenger; C402487:RPL13A protein, human; D019707:Receptors, Chemokine; D012269:Ribosomal Proteins; D007371:Interferon-gamma; D002570:Ceruloplasmin