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Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA
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 whethe...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5513066/ https://www.ncbi.nlm.nih.gov/pubmed/28455422 http://dx.doi.org/10.1261/rna.060467.116 |
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author | Ball, Christopher B. Solem, Amanda C. Meganck, Rita M. Laederach, Alain Ramos, Silvia B.V. |
author_facet | Ball, Christopher B. Solem, Amanda C. Meganck, Rita M. Laederach, Alain Ramos, Silvia B.V. |
author_sort | Ball, Christopher B. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5513066 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-55130662018-08-01 Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA Ball, Christopher B. Solem, Amanda C. Meganck, Rita M. Laederach, Alain Ramos, Silvia B.V. RNA Article 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. Cold Spring Harbor Laboratory Press 2017-08 /pmc/articles/PMC5513066/ /pubmed/28455422 http://dx.doi.org/10.1261/rna.060467.116 Text en © 2017 Ball et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Article Ball, Christopher B. Solem, Amanda C. Meganck, Rita M. Laederach, Alain Ramos, Silvia B.V. Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title_full | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title_fullStr | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title_full_unstemmed | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title_short | Impact of RNA structure on ZFP36L2 interaction with luteinizing hormone receptor mRNA |
title_sort | impact of rna structure on zfp36l2 interaction with luteinizing hormone receptor mrna |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5513066/ https://www.ncbi.nlm.nih.gov/pubmed/28455422 http://dx.doi.org/10.1261/rna.060467.116 |
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