Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction
The RNA binding protein heterogeneous nuclear ribonucleoprotein A1 (A1) regulates RNA metabolism, which is crucial to maintaining cellular homeostasis. A1 dysfunction mechanistically contributes to reduced cell viability and loss, but molecular mechanisms of how A1 dysfunction affects cell viability...
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| Format: | Article |
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Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Molecular Biosciences |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1178439/full |
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| author | Joseph P. Clarke Joseph P. Clarke Patricia A. Thibault Patricia A. Thibault Sakina Fatima Hannah E. Salapa Hannah E. Salapa Subha Kalyaanamoorthy Aravindhan Ganesan Michael C. Levin Michael C. Levin Michael C. Levin Michael C. Levin |
| author_facet | Joseph P. Clarke Joseph P. Clarke Patricia A. Thibault Patricia A. Thibault Sakina Fatima Hannah E. Salapa Hannah E. Salapa Subha Kalyaanamoorthy Aravindhan Ganesan Michael C. Levin Michael C. Levin Michael C. Levin Michael C. Levin |
| author_sort | Joseph P. Clarke |
| collection | DOAJ |
| description | The RNA binding protein heterogeneous nuclear ribonucleoprotein A1 (A1) regulates RNA metabolism, which is crucial to maintaining cellular homeostasis. A1 dysfunction mechanistically contributes to reduced cell viability and loss, but molecular mechanisms of how A1 dysfunction affects cell viability and loss, and methodologies to attenuate its dysfunction, are lacking. Utilizing in silico molecular modeling and an in vitro optogenetic system, this study examined the consequences of RNA oligonucleotide (RNAO) treatment on attenuating A1 dysfunction and its downstream cellular effects. In silico and thermal shift experiments revealed that binding of RNAOs to the RNA Recognition Motif 1 of A1 is stabilized by sequence- and structure-specific RNAO-A1 interactions. Using optogenetics to model A1 cellular dysfunction, we show that sequence- and structure-specific RNAOs significantly attenuated abnormal cytoplasmic A1 self-association kinetics and A1 cytoplasmic clustering. Downstream of A1 dysfunction, we demonstrate that A1 clustering affects the formation of stress granules, activates cell stress, and inhibits protein translation. With RNAO treatment, we show that stress granule formation is attenuated, cell stress is inhibited, and protein translation is restored. This study provides evidence that sequence- and structure-specific RNAO treatment attenuates A1 dysfunction and its downstream effects, thus allowing for the development of A1-specific therapies that attenuate A1 dysfunction and restore cellular homeostasis. |
| first_indexed | 2024-03-13T03:53:21Z |
| format | Article |
| id | doaj.art-e9a6b9cc5b554f6ca0e0e99458443344 |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-03-13T03:53:21Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-e9a6b9cc5b554f6ca0e0e994584433442023-06-22T09:33:47ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2023-06-011010.3389/fmolb.2023.11784391178439Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunctionJoseph P. Clarke0Joseph P. Clarke1Patricia A. Thibault2Patricia A. Thibault3Sakina Fatima4Hannah E. Salapa5Hannah E. Salapa6Subha Kalyaanamoorthy7Aravindhan Ganesan8Michael C. Levin9Michael C. Levin10Michael C. Levin11Michael C. Levin12Department of Health Sciences, College of Medicine, University of Saskatchewan, Saskatoon, SK, CanadaOffice of the Saskatchewan Multiple Sclerosis Clinical Research Chair, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Medicine, Neurology Division, University of Saskatchewan, Saskatoon, SK, CanadaOffice of the Saskatchewan Multiple Sclerosis Clinical Research Chair, University of Saskatchewan, Saskatoon, SK, CanadaArGan’s Lab, School of Pharmacy, Faculty of Science, University of Waterloo, Waterloo, ON, CanadaDepartment of Medicine, Neurology Division, University of Saskatchewan, Saskatoon, SK, CanadaOffice of the Saskatchewan Multiple Sclerosis Clinical Research Chair, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Chemistry, Faculty of Science, University of Waterloo, Waterloo, ON, CanadaArGan’s Lab, School of Pharmacy, Faculty of Science, University of Waterloo, Waterloo, ON, CanadaDepartment of Health Sciences, College of Medicine, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Medicine, Neurology Division, University of Saskatchewan, Saskatoon, SK, CanadaDepartment of Anatomy, Physiology and Pharmacology, University of Saskatchewan, Saskatoon, SK, CanadaOffice of the Saskatchewan Multiple Sclerosis Clinical Research Chair, University of Saskatchewan, Saskatoon, SK, CanadaThe RNA binding protein heterogeneous nuclear ribonucleoprotein A1 (A1) regulates RNA metabolism, which is crucial to maintaining cellular homeostasis. A1 dysfunction mechanistically contributes to reduced cell viability and loss, but molecular mechanisms of how A1 dysfunction affects cell viability and loss, and methodologies to attenuate its dysfunction, are lacking. Utilizing in silico molecular modeling and an in vitro optogenetic system, this study examined the consequences of RNA oligonucleotide (RNAO) treatment on attenuating A1 dysfunction and its downstream cellular effects. In silico and thermal shift experiments revealed that binding of RNAOs to the RNA Recognition Motif 1 of A1 is stabilized by sequence- and structure-specific RNAO-A1 interactions. Using optogenetics to model A1 cellular dysfunction, we show that sequence- and structure-specific RNAOs significantly attenuated abnormal cytoplasmic A1 self-association kinetics and A1 cytoplasmic clustering. Downstream of A1 dysfunction, we demonstrate that A1 clustering affects the formation of stress granules, activates cell stress, and inhibits protein translation. With RNAO treatment, we show that stress granule formation is attenuated, cell stress is inhibited, and protein translation is restored. This study provides evidence that sequence- and structure-specific RNAO treatment attenuates A1 dysfunction and its downstream effects, thus allowing for the development of A1-specific therapies that attenuate A1 dysfunction and restore cellular homeostasis.https://www.frontiersin.org/articles/10.3389/fmolb.2023.1178439/fullhnRNPA1RNA oligonucleotideoptogeneticsstress granulesRNA-protein interaction |
| spellingShingle | Joseph P. Clarke Joseph P. Clarke Patricia A. Thibault Patricia A. Thibault Sakina Fatima Hannah E. Salapa Hannah E. Salapa Subha Kalyaanamoorthy Aravindhan Ganesan Michael C. Levin Michael C. Levin Michael C. Levin Michael C. Levin Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction Frontiers in Molecular Biosciences hnRNPA1 RNA oligonucleotide optogenetics stress granules RNA-protein interaction |
| title | Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction |
| title_full | Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction |
| title_fullStr | Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction |
| title_full_unstemmed | Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction |
| title_short | Sequence- and structure-specific RNA oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein A1 dysfunction |
| title_sort | sequence and structure specific rna oligonucleotide binding attenuates heterogeneous nuclear ribonucleoprotein a1 dysfunction |
| topic | hnRNPA1 RNA oligonucleotide optogenetics stress granules RNA-protein interaction |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2023.1178439/full |
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