Abstract
Replication-dependent histone (RDH) mRNAs have a nonpolyadenylated 3'-UTR that ends in a highly conserved stem-loop structure. Nonetheless, a subset of RDH mRNAs has a poly(A) tail under physiological conditions. The biological meaning of poly(A)-containing (+) RDH mRNAs and details of their biosynthesis remain elusive. Here, using HeLa cells and Western blotting, qRT-PCR, and biotinylated RNA pulldown assays, we show that poly(A)+ RDH mRNAs are post-transcriptionally regulated via adenylate- and uridylate-rich element-mediated mRNA decay (AMD). We observed that the rapid degradation of poly(A)+ RDH mRNA is driven by butyrate response factor 1 (BRF1; also known as ZFP36 ring finger protein-like 1) under normal conditions. Conversely, cellular stresses such as UV C irradiation promoted BRF1 degradation, increased the association of Hu antigen R (HuR; also known as ELAV-like RNA-binding protein 1) with the 3'-UTR of poly(A)+ RDH mRNAs, and eventually stabilized the poly(A)+ RDH mRNAs. Collectively, our results provide evidence that AMD surveils poly(A)+ RDH mRNAs via BRF1-mediated degradation under physiological conditions.
Highlights
Replication-dependent histone (RDH) mRNAs have a nonpolyadenylated 3-UTR that ends in a highly conserved stemloop structure
The extended 3Ј-UTR of poly(A)ϩ RDH mRNAs may contain a new cis-acting element(s), which can serve as a platform for a different regulatory mode of gene expression, as compared with HSLϩ RDH mRNAs
We provide evidence that butyrate response factor 1 (BRF1) promotes rapid degradation of poly(A)ϩ RDH mRNAs via an association between BRF1 and AREs within the PRSE in the extended 3Ј-UTRs
Summary
Inhibition of proteasomes abrogates stress-mediated stabilization of poly(A)؉ RDH mRNAs. The up-regulation of HIST1H4KMT10 mRNA upon Puro treatment was reversed by MG132 treatment (Fig. 1, D and F) All these data suggest that stabilization of poly(A)ϩ RDH mRNA via a PRSE under stressful conditions depends on a proteasomal activity and that a putative labile cellular factor(s) is responsible for rapid degradation of poly(A)ϩ RDH mRNAs under normal conditions. We determined whether BRFs bind to poly(A)ϩ RDH mRNA and whether their interaction is affected by cellular stresses To this end, we performed a pulldown assay using in vitro–synthesized biotinylated RNA, either PRSE RNA or PRSE-Rev RNA, and the extracts obtained from UVC-irradiated or nonirradiated HeLa cells (Fig. 3A). Cate that BRF1 is a bona fide decay-promoting factor for poly(A)ϩ RDH mRNAs
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