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RNA-Seq analysis of wild-type and ttpA-C231S, ttpA-E236fsx, and ttpA-Y242fsx mutant D. discoideum amebae

ABSTRACT: In many eukaryotes, mRNAs containing specific AU-rich motifs are regulated by proteins of the tristetraprolin (TTP) family, which bind to these motifs through a tandem zinc finger (TZF) domain. This binding leads to promotion of subsequent deadenylation and decay, partly through a conserved carboxyl-terminal CNOT1 binding domain. We explored the physiological role of the single TTP family member (TtpA) expressed in Dictyostelium discoideum. This study shows the effect of three distinct putative null mutations in the ttpA gene on transcript expression in amebae in growth medium during surface culture under axenic conditions. The cells exhibited similar external and gene expression phenotypes, suggesting that they are all null mutants, despite two different types of mutations (two frame shifts and one amino acid substitution).

ORGANISM(S): Dictyostelium discoideum

PROVIDER: GSE156809 | GEO | 2021/11/24

REPOSITORIES: GEO

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RNA-Seq analysis of wild-type and ttpA-D450trx mutant D. discoideum amebae

Project description:In many eukaryotes, mRNAs containing specific AU-rich motifs are regulated by proteins of the tristetraprolin (TTP) family, which bind to these motifs through a tandem zinc finger (TZF) domain. This binding leads to promotion of subsequent deadenylation and decay, partly through a conserved carboxyl-terminal CNOT1 binding domain. We explored the physiological role of the single TTP family member (TtpA) expressed in Dictyostelium discoideum. This study shows the effect of a carboxyl-terminal truncation (ttpA-D450trx), which removed the predicted CNOT1 binding domain, evaluated in amebae in growth medium during surface culture. The cells exhibited external and gene expression phenotypes that were very similar to those of two distinct null mutants, raising the possibility that this domain is necessary for TtpA-promoted mRNA decay in this species.

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TTP-dependent mRNA decay in LPS-stimulated macrophages

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RNA-Seq analysis of wild-type and ttpA-D450trx mutant D. discoideum amebae

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Posttranscriptional control of the neutrophil transcriptome by tristetraprolin promotes neutrophil apoptosis and compromises host antimicrobial defense

Project description:Posttranscriptional regulation of mRNA levels in neutrophils and its consequences for immune responses are unexplored. By employing profiling of the neutrophil transcriptome we show that the mRNA-destabilizing protein tristetraprolin (TTP) limits the expression of hundreds of genes, including genes negatively regulating apoptosis. Elicited TTP-deficient neutrophils exhibited reduced apoptosis and were increased in numbers. The anti-apoptotic protein Mcl-1 was elevated in TTP-deficient neutrophils and Mcl1 mRNA was bound and destabilized by TTP. Ablation of TTP in macrophages and neutrophils resulted in an improved defense and survival of mice during invasive infection with Streptococcus pyogenes. Mice lacking myeloid TTP prevented dissemination of bacteria and efficiently blunted systemic disease by massive but controlled neutrophil deployment. These data identify posttranscriptional control by TTP to restrict neutrophils and antimicrobial defense.

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Project description:Controlled decay of cytokine and chemokine mRNAs restrains the time and amplitude of inflammatory responses. Tristetraprolin (TTP) binds to AU-rich elements in 3´ untranslated regions of mRNA and targets the bound mRNA for degradation. We have addressed here the function of TTP in balancing the macrophage activation state by a comprehensive analysis of TTP-dependent mRNA decay in LPS-stimulated macrophages from WT and TTP-deficient mice. We compared mRNA stability in LPS-treated BMDMs from WT and TTP-/- mice by microarray-based measurement of the remnant mRNA after transcriptional blockade with actinomycin D (act D). To increase the sensitivity of the mRNA decay profiling we inhibited the LPS-activated p38 MAPK with the specific inhibitor SB203580 since p38 MAPK negatively regulates the mRNA-destabilizing activity of TTP. LPS stimulation was for 3h before addition of act D. RNA was harvested at 0', 45' and 90' thereafter.

2011-12-15 | E-GEOD-28880 | biostudies-arrayexpress

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