Tristetraprolin regulates CXCL1 (KC) mRNA stability
Shyamasree Datta1, Roopa Biswas, Michael Novotny
1Department of Immunology, Lerner Research Institute, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
Journal of Immunology (Baltimore, Md. : 1950)
|February 6, 2008
Summary
Tristetraprolin (TTP) is the main regulator of chemokine KC mRNA decay in immune cells, targeting specific sequences in the mRNA to control its stability and inflammatory responses.
Area of Science:
- Molecular Biology
- Immunology
- Gene Regulation
Background:
- Proinflammatory chemokine mRNA stability is controlled by adenine-uridine-rich sequences (AREs) in the 3' untranslated region.
- Tristetraprolin (TTP) is an RNA-binding protein known to regulate the decay of several ARE-containing mRNAs.
Purpose of the Study:
- To investigate the role of TTP in regulating the decay of mouse chemokine KC (CXCL1) mRNA.
- To identify sequence determinants and cellular contexts governing TTP-mediated mRNA decay.
Main Methods:
- Utilized TTP-deficient HEK293 cells with Tet-On system for transcriptional control.
- Employed deletion and site-specific mutagenesis to map AREs.
- Compared mRNA decay in primary macrophages from wild-type and TTP-deficient mice.
- Assessed TTP-mediated decay sensitivity to IL-1alpha and LPS stimulation, including p38 MAPK inhibition.
Main Results:
- TTP significantly decreased KC mRNA half-life in HEK293 cells.
- Multiple AUUUA motifs within the 3' UTR were identified as critical for TTP sensitivity.
- KC mRNA instability in macrophages was highly TTP-dependent.
- LPS-induced KC mRNA stabilization in wild-type macrophages was abrogated by p38 MAPK inhibition, but not in TTP-deficient cells.
- TTP-mediated decay in 293 cells was insensitive to IL-1alpha stimulation.
Conclusions:
- TTP is the predominant regulator of KC mRNA decay in mononuclear phagocytes, acting through multiple 3'-UTR AREs.
- Additional factors contribute to KC mRNA instability and stimulus sensitivity in TTP-independent cellular contexts.
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