Genome-wide analysis identifies interleukin-10 mRNA as target of tristetraprolin.
Tristetraprolin (TTP) is an RNA-binding protein required for the rapid degradation of mRNAs containing AU-rich elements. Targets regulated by TTP include the mRNAs encoding tumor necrosis factor-alpha, granulocyte-macrophage colony-stimulating factor, interleukin-2 (IL-2), and immediate early response 3. To identify novel target mRNAs of TTP in macrophages, we used a genome-wide approach that combines RNA immunoprecipitation and microarray analysis. A list was compiled of 137 mRNAs that are associated with TTP with an estimated accuracy on the order of 90%. Sequence analysis revealed a highly significant enrichment of AU-rich element motifs, with AUUUA pentamers present in 96% and UUAUUUAUU nonamers present in 44% of TTP-associated mRNAs. We further show that IL-10 is a novel target regulated by TTP. IL-10 mRNA levels were found to be elevated because of a reduced decay rate in primary macrophages from TTP(-/-) mice. Our study demonstrates the importance of experimental approaches for identifying targets of RNA-binding proteins.
Stoecklin
Georg
G
Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA. g.stoecklin@dkfz.de
Tenenbaum
Scott A
SA
Mayo
Thomas
T
Chittur
Sridar V
SV
George
Ajish D
AD
Baroni
Timothy E
TE
Blackshear
Perry J
PJ
Anderson
Paul
P
D020413:3' Untranslated Regions; D000818:Animals; D002460:Cell Line; D005786:Gene Expression Regulation; D016678:Genome; D007249:Inflammation; D016753:Interleukin-10 / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis; D017434:Protein Structure, Tertiary; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism; D051816:Tristetraprolin / Q000378:metabolism*
D020413:3' Untranslated Regions; D012333:RNA, Messenger; D051816:Tristetraprolin; D016753:Interleukin-10