Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN
The 24‐h molecular clock is based on the stability of rhythmically expressed transcripts. The shortening of the poly(A) tail of mRNAs is often the first and rate‐limiting step that determines the lifespan of a mRNA and is catalyzed by deadenylases. Herein, we determine the catalytic site of Hesperin...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-05-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.13011 |
| _version_ | 1828221576270577664 |
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| author | Rafailia A. A. Beta Athanasios Kyritsis Veroniki Douka Eirini Papanastasi Marianna Rizouli Demetres D. Leonidas Dimitrios Vlachakis Nikolaos A. A. Balatsos |
| author_facet | Rafailia A. A. Beta Athanasios Kyritsis Veroniki Douka Eirini Papanastasi Marianna Rizouli Demetres D. Leonidas Dimitrios Vlachakis Nikolaos A. A. Balatsos |
| author_sort | Rafailia A. A. Beta |
| collection | DOAJ |
| description | The 24‐h molecular clock is based on the stability of rhythmically expressed transcripts. The shortening of the poly(A) tail of mRNAs is often the first and rate‐limiting step that determines the lifespan of a mRNA and is catalyzed by deadenylases. Herein, we determine the catalytic site of Hesperin, a recently described circadian deadenylase in plants, using a modified site‐directed mutagenesis protocol and a custom vector, pATHRA. To explore the catalytic efficiency of AtHESPERIN, we investigated the effect of AMP and neomycin, and used molecular modeling simulations to propose a catalytic mechanism. Collectively, the biochemical and in silico results classify AtHESPERIN in the exonuclease–endonuclease–phosphatase deadenylase superfamily and contribute to the understanding of the intricate mechanisms of circadian mRNA turnover. |
| first_indexed | 2024-04-12T16:43:28Z |
| format | Article |
| id | doaj.art-96c2b72e4e1b4f3cb35595297c948d75 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-12T16:43:28Z |
| publishDate | 2022-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-96c2b72e4e1b4f3cb35595297c948d752022-12-22T03:24:42ZengWileyFEBS Open Bio2211-54632022-05-011251036104910.1002/2211-5463.13011Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERINRafailia A. A. Beta0Athanasios Kyritsis1Veroniki Douka2Eirini Papanastasi3Marianna Rizouli4Demetres D. Leonidas5Dimitrios Vlachakis6Nikolaos A. A. Balatsos7Department of Biochemistry and Biotechnology University of Thessaly Larissa GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceGenetics Laboratory Department of Biotechnology Agricultural University of Athens GreeceDepartment of Biochemistry and Biotechnology University of Thessaly Larissa GreeceThe 24‐h molecular clock is based on the stability of rhythmically expressed transcripts. The shortening of the poly(A) tail of mRNAs is often the first and rate‐limiting step that determines the lifespan of a mRNA and is catalyzed by deadenylases. Herein, we determine the catalytic site of Hesperin, a recently described circadian deadenylase in plants, using a modified site‐directed mutagenesis protocol and a custom vector, pATHRA. To explore the catalytic efficiency of AtHESPERIN, we investigated the effect of AMP and neomycin, and used molecular modeling simulations to propose a catalytic mechanism. Collectively, the biochemical and in silico results classify AtHESPERIN in the exonuclease–endonuclease–phosphatase deadenylase superfamily and contribute to the understanding of the intricate mechanisms of circadian mRNA turnover.https://doi.org/10.1002/2211-5463.13011AtHESPERINcircadian rhythmsdeadenylationmRNA decaypoly(A) tail |
| spellingShingle | Rafailia A. A. Beta Athanasios Kyritsis Veroniki Douka Eirini Papanastasi Marianna Rizouli Demetres D. Leonidas Dimitrios Vlachakis Nikolaos A. A. Balatsos Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN FEBS Open Bio AtHESPERIN circadian rhythms deadenylation mRNA decay poly(A) tail |
| title | Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN |
| title_full | Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN |
| title_fullStr | Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN |
| title_full_unstemmed | Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN |
| title_short | Biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase HESPERIN |
| title_sort | biochemical and in silico identification of the active site and the catalytic mechanism of the circadian deadenylase hesperin |
| topic | AtHESPERIN circadian rhythms deadenylation mRNA decay poly(A) tail |
| url | https://doi.org/10.1002/2211-5463.13011 |
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