[PMID]: | 28455422 |
[Au] Autor: | Ball CB; Solem AC; Meganck RM; Laederach A; Ramos SBV |
[Ad] Endereço: | Biochemistry and Biophysics Department, University of North Carolina, Chapel Hill, North Carolina 27599, USA. |
[Ti] Título: | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA. |
[So] Source: | RNA;23(8):1209-1223, 2017 08. |
[Is] ISSN: | 1469-9001 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | ZFP36L2 (L2) destabilizes AU-rich element (ARE)-containing transcripts and has been implicated in female fertility. We have shown that only one of three putative AREs within the 3' UTR of murine luteinizing hormone receptor mRNA, ARE2197 (UAUUUAU), is capable of interacting with L2. To assess whether structural elements of ARE2197 could explain this unique binding ability, we performed whole-transcript SHAPE-MaP (selective 2' hydroxyl acylation by primer extension-mutational profiling) of the full-length mLHR mRNA. The data revealed that the functional ARE2197 is located in a hairpin loop structure and most nucleotides are highly reactive. In contrast, each of the nonbinding AREs, 2301 and 2444, contains only a pentamer AUUUA; and in ARE2301 much of the ARE sequence is poorly accessible. Because the functional mARE was also found to be conserved in humans at the sequence level (ARE 2223), we decided to investigate whether binding and structure are also preserved. Similar to mouse, only one ARE in hLHR mRNA is capable of binding to L2; and it is also located in a hairpin structure, based on our SHAPE-MaP data. To investigate the role of secondary structure in the binding, we mutated specific nucleotides in both functional AREs. Mutations in the flexible stem region proximal to the loop that enforce strong base-pairing, drastically reduced L2 binding affinity; this confirms that the structural context is critical for L2 recognition of hARE2223. Collectively, our results suggest that a combination of minimal ARE sequence, placement of the ARE in a hairpin loop, and stem flexibility mediate high-affinity L2 binding to hLHR mRNA. |
[Mh] Termos MeSH primário: |
Elementos Ricos em Adenilato e Uridilato/genética RNA Mensageiro/metabolismo Receptores do LH/metabolismo Tristetraprolina/metabolismo
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[Mh] Termos MeSH secundário: |
Animais Pareamento de Bases Sequência de Bases Seres Humanos Camundongos Mutação/genética Conformação de Ácido Nucleico RNA Mensageiro/química RNA Mensageiro/genética Receptores do LH/genética Alinhamento de Sequência Tristetraprolina/química Tristetraprolina/genética
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[Pt] Tipo de publicação: | JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T; RESEARCH SUPPORT, N.I.H., EXTRAMURAL |
[Nm] Nome de substância:
| 0 (RNA, Messenger); 0 (Receptors, LH); 0 (Tristetraprolin); 0 (Zfp36 protein, mouse) |
[Em] Mês de entrada: | 1709 |
[Cu] Atualização por classe: | 171223 |
[Lr] Data última revisão:
| 171223 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 170430 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1261/rna.060467.116 |
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