In vitro characterization of the yeast DEAH/RHA RNA helicase Dhr1.
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| Title: | In vitro characterization of the yeast DEAH/RHA RNA helicase Dhr1. |
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| Authors: | Lin, Ran1, Elmir, Ezzeddine2, Reynolds, Madison J.3, Johnson, Arlen W.1 arlen@utexas.edu |
| Source: | Journal of Biological Chemistry. Apr2025, Vol. 301 Issue 4, p1-13. 13p. |
| Subjects: | RNA helicase, Helicases, Organelle formation, Ribosomal RNA, Adenosine triphosphatase |
| Abstract: | In eukaryotic ribosome biogenesis, the small subunit (SSU) processome is a metastable intermediate in the assembly of the small (40S) subunit. In the SSU processome, the ribosomal RNA domains are splayed open by the intervention of assembly factors as well as U3 snoRNA. A critical step during the transition from the SSU processome to the nearly mature pre-40S particle is the removal of the U3 snoRNA to allow the formation of the central pseudoknot, a universally conserved structure which connects all domains of the subunit and contributes to its dynamic nature during translation. We previously identified the DEAH/RHA RNA helicase Dhr1 as the enzyme responsible for displacing the U3 snoRNA and the SSU processome factor Utp14 as an activator of Dhr1. Here, we have utilized biochemical and yeast genetic methods to further characterize Dhr1. We show that the N terminus as well as an internal loop within the RecA2 domain are autoinhibitory. We found that Utp14 can activate the ATPase activity of Dhr1 lacking the autoinhibitory N-terminal loop but not full-length Dhr1. We considered the possibility that Utp14 activates Dhr1 by relieving the autoinhibition of the loop within the RecA2 domain. However, our results are more consistent with Utp14 activating Dhr1 by binding to the surface of the RecA1 and RecA2 domains rather than displacing the inhibitory loop. This position of Utp14 is distinct from how G-patch proteins activate other DEXH/RHA helicases and is consistent with our previous conclusion that Utp14 is not a canonical G-patch protein. [ABSTRACT FROM AUTHOR] |
| Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
| FullText | Text: Availability: 0 |
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| Header | DbId: egs DbLabel: Engineering Source An: 185344949 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: In vitro characterization of the yeast DEAH/RHA RNA helicase Dhr1. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Lin%2C+Ran%22">Lin, Ran</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Elmir%2C+Ezzeddine%22">Elmir, Ezzeddine</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Reynolds%2C+Madison+J%2E%22">Reynolds, Madison J.</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Johnson%2C+Arlen+W%2E%22">Johnson, Arlen W.</searchLink><relatesTo>1</relatesTo><i> arlen@utexas.edu</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+Biological+Chemistry%22">Journal of Biological Chemistry</searchLink>. Apr2025, Vol. 301 Issue 4, p1-13. 13p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22RNA+helicase%22">RNA helicase</searchLink><br /><searchLink fieldCode="DE" term="%22Helicases%22">Helicases</searchLink><br /><searchLink fieldCode="DE" term="%22Organelle+formation%22">Organelle formation</searchLink><br /><searchLink fieldCode="DE" term="%22Ribosomal+RNA%22">Ribosomal RNA</searchLink><br /><searchLink fieldCode="DE" term="%22Adenosine+triphosphatase%22">Adenosine triphosphatase</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: In eukaryotic ribosome biogenesis, the small subunit (SSU) processome is a metastable intermediate in the assembly of the small (40S) subunit. In the SSU processome, the ribosomal RNA domains are splayed open by the intervention of assembly factors as well as U3 snoRNA. A critical step during the transition from the SSU processome to the nearly mature pre-40S particle is the removal of the U3 snoRNA to allow the formation of the central pseudoknot, a universally conserved structure which connects all domains of the subunit and contributes to its dynamic nature during translation. We previously identified the DEAH/RHA RNA helicase Dhr1 as the enzyme responsible for displacing the U3 snoRNA and the SSU processome factor Utp14 as an activator of Dhr1. Here, we have utilized biochemical and yeast genetic methods to further characterize Dhr1. We show that the N terminus as well as an internal loop within the RecA2 domain are autoinhibitory. We found that Utp14 can activate the ATPase activity of Dhr1 lacking the autoinhibitory N-terminal loop but not full-length Dhr1. We considered the possibility that Utp14 activates Dhr1 by relieving the autoinhibition of the loop within the RecA2 domain. However, our results are more consistent with Utp14 activating Dhr1 by binding to the surface of the RecA1 and RecA2 domains rather than displacing the inhibitory loop. This position of Utp14 is distinct from how G-patch proteins activate other DEXH/RHA helicases and is consistent with our previous conclusion that Utp14 is not a canonical G-patch protein. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1016/j.jbc.2025.108366 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 13 StartPage: 1 Subjects: – SubjectFull: RNA helicase Type: general – SubjectFull: Helicases Type: general – SubjectFull: Organelle formation Type: general – SubjectFull: Ribosomal RNA Type: general – SubjectFull: Adenosine triphosphatase Type: general Titles: – TitleFull: In vitro characterization of the yeast DEAH/RHA RNA helicase Dhr1. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Lin, Ran – PersonEntity: Name: NameFull: Elmir, Ezzeddine – PersonEntity: Name: NameFull: Reynolds, Madison J. – PersonEntity: Name: NameFull: Johnson, Arlen W. IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 04 Text: Apr2025 Type: published Y: 2025 Identifiers: – Type: issn-print Value: 00219258 Numbering: – Type: volume Value: 301 – Type: issue Value: 4 Titles: – TitleFull: Journal of Biological Chemistry Type: main |
| ResultId | 1 |
