Sequence and tissue targeting specificity of ZFP36L2 reveals Elavl2 as a novel target with co-regulation potential

Abstract

Zinc finger protein 36 like 2 (ZFP36L2) is an RNA-binding protein that destabilizes transcripts containing adenine-uridine rich elements (AREs). The overlap between ZFP36L2 targets in different tissues is minimal, suggesting that ZFP36L2-targeting is highly tissue specific. We developed a novel Zfp36l2-lacking mouse model (L2-fKO) to identify factors governing this tissue specificity. We found 549 upregulated genes in the L2-fKO spleen by RNA-seq. These upregulated genes were enriched in ARE motifs in the 3′UTRs, which suggests that they are ZFP36L2 targets, however the precise sequence requirement for targeting was not evident from motif analysis alone. We therefore used gel-shift mobility assays on 12 novel putative targets and established that ZFP36L2 requires a 7-mer (UAUUUAU) motif to bind. We observed a statistically significant enrichment of 7-mer ARE motifs in upregulated genes and determined that ZFP36L2 targets are enriched for multiple 7-mer motifs. Elavl2 mRNA, which has three 7-mer (UAUUUAU) motifs, was also upregulated in L2-fKO spleens. Overexpression of ZFP36L2, but not a ZFP36L2(C176S) mutant, reduced Elavl2 mRNA expression, suggesting a direct negative effect. Additionally, a reporter assay demonstrated that the ZFP36L2 effect on Elavl2 decay is dependent on the Elavl2-3′UTR and requires the 7-mer AREs. Our data indicate that Elavl2 mRNA is a novel target of ZFP36L2, specific to the spleen. Likely, ZFP36L2 combined with other RNA binding proteins, such as ELAVL2, governs tissue specificity.

Structural Modifications and Novel Protein-Binding Sites in Pre-miR-675—Explaining Its Regulatory Mechanism in Carcinogenesis

“…The overexpression of Lin28b is linked with the development of ovarian cancer [ 32 ] and cellular proliferation in acute myeloid leukaemia [ 33 ]. Likewise, HuR is an RNA-binding protein belonging to the embryonic lethal and altered vision (ELAV) family of proteins [ 34 ] and is essential for the stability and translation of mRNAs [ 23 ]. Interactions between HuR and other miRNAs have been implicated in the development and progression of various cancers [ 35 , 36 , 37 ].…”

Transcriptomic and epigenomic landscapes of muscle growth during the postnatal period of broilers

“…Differentially expressed genes (DEGs) between different groups were identified by DESeq2 package (v.1.36.0) [ 29 ] in the R program. Genes with an adjusted P value < 0.05 and a log 2 (Fold Change) > |1| were considered significant DEGs [ 30 – 32 ]. We referred to the DEGs that were up-regulated in CC among the breed-biased genes as CC-biased DEGs (CBGs).…”