Intact NYN/PIN‐Like Domain is Crucial for the Degradation of Inflammation‐Related Transcripts by ZC3H12D

ABSTRACT ZC3H12D belongs to a recently discovered family of proteins containing four members of which the most studied and best described is the RNase ZC3H12A (MCPIP1/Regnase‐1). ZC3H12A is a crucial negative regulator of inflammation. It accelerates the turnover of transcripts of a spectrum of proinflammatory cytokines, as well as its own mRNA. The biological role of ZC3H12D is less clear, although it was shown that this member of ZC3H12 family is also involved in the regulation of inflammation. Here, we show that ZC3H12A and ZC3H12D recognize a set of common target mRNAs encoding proteins that play important roles in the course of the inflammation. Similarly to ZC3H12A, ZC3H12D participates in the 3′UTR‐dependent regulation of the turnover of mRNAs encoding interleukin‐6 (IL‐6), tumor necrosis factor (TNF), and immediate early response 3 gene (IER3). The ZC3H12A mRNA is also among the identified ZC3H12D targets. Using the combination of immunofluorescence with single molecule RNA fluorescence in situ hybridization (smRNA FISH) we have shown that ZC3H12D protein interacts with the ZC3H12A transcript. The direct binding of these two molecules in vivo was further confirmed by RNA immunoprecipitation. Simultaneously, overexpression of ZC3H12D increases the turnover rate of transcripts containing ZC3H12A 3′UTR. Using reporter gene assays we have confirmed that the Asp95 residue present in the NYN/PIN‐like domain is crucial for ZC3H12D biological activity. We have also revealed that ZC3H12D recognizes the same structural elements present in the 3′UTRs of the investigated transcripts, as ZC3H12A. J. Cell. Biochem. 118: 487–498, 2017. © 2016 Wiley Periodicals, Inc. Protein ZC3H12D regulates the half‐life of transcripts in a NYN/PIN‐like domain‐dependent manner. Besides IL‐6, TNF, and IER3 mRNAs, ZC3H12D interacts also with the transcript of ZC3H12A (MCPIP1), a known inflammation regulator.