Subunit D of RNA Polymerase from Methanosarcina acetivorans Contains Two Oxygen-labile [4Fe-4S] Clusters: IMPLICATIONS FOR OXIDANT-DEPENDENT REGULATION OF TRANSCRIPTION.
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| Title: | Subunit D of RNA Polymerase from Methanosarcina acetivorans Contains Two Oxygen-labile [4Fe-4S] Clusters: IMPLICATIONS FOR OXIDANT-DEPENDENT REGULATION OF TRANSCRIPTION. |
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| Authors: | Lessner, Faith H.1, Jennings, Matthew E.1, Hirata, Akira2, Duin, Eduardus C.3, Lessner, Daniel J.1 dlessner@uark.edu. |
| Source: | Journal of Biological Chemistry. 5/25/2012, Vol. 287 Issue 22, p18510-18523. 14p. |
| Subjects: | RNA polymerases, Methanosarcina, Genomes, Escherichia coli, Transcription factors |
| Abstract: | Subunit D of multisubunit RNA polymerase from many species of archaea is predicted to bind one to two iron-sulfur (Fe-S) clusters, the function of which is unknown. A survey of encoded subunit D in the genomes of sequenced archaea revealed six distinct groups based on the number of complete or partial [4Fe- 4S] cluster motifs within domain 3. Only subunitDfrom strictly anaerobic archaea, including all members of the Methanosarcinales, are predicted to bind two [4Fe-4S] clusters. We report herein the purification and characterization of Methanosarcina acetivorans subunit D in complex with subunit L. Expression of subunit D and subunit L in Escherichia coli resulted in the purification of a D-L heterodimer with only partial [4Fe-4S] cluster content. Reconstitution in vitro with iron and sulfide revealed that the M. acetivorans D-L heterodimer is capable of binding two redox-active [4Fe-4S] clusters. M. acetivorans subunit D deleted of domain 3 (DΔD3) was still capable of co-purifying with subunit L but was devoid of [4Fe-4S] clusters. Affinity purification of subunit D or subunit DΔD3 from M. acetivorans resulted in the co-purification of endogenous subunit L with each tagged subunit D. Overall, these results suggest that domain 3 of subunit D is required for [4Fe-4S] cluster binding, but the [4Fe-4S] clusters and domain 3 are not required for the formation of the D-L heterodimer. However, exposure of two [4Fe-4S] cluster-containing D-L heterodimer to oxygen resulted in loss of the [4Fe-4S] clusters and subsequent protein aggregation, indicating that the [4Fe-4S] clusters influence the stability of the D-L heterodimer and therefore have the potential to regulate the assembly and/or activity of RNA polymerase in an oxidant- dependent manner. [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 |
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| Header | DbId: egs DbLabel: Engineering Source An: 76568160 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Subunit D of RNA Polymerase from Methanosarcina acetivorans Contains Two Oxygen-labile [4Fe-4S] Clusters: IMPLICATIONS FOR OXIDANT-DEPENDENT REGULATION OF TRANSCRIPTION. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Lessner%2C+Faith+H%2E%22">Lessner, Faith H.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Jennings%2C+Matthew+E%2E%22">Jennings, Matthew E.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Hirata%2C+Akira%22">Hirata, Akira</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Duin%2C+Eduardus+C%2E%22">Duin, Eduardus C.</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Lessner%2C+Daniel+J%2E%22">Lessner, Daniel J.</searchLink><relatesTo>1</relatesTo><i> dlessner@uark.edu.</i> – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Journal+of+Biological+Chemistry%22">Journal of Biological Chemistry</searchLink>. 5/25/2012, Vol. 287 Issue 22, p18510-18523. 14p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22RNA+polymerases%22">RNA polymerases</searchLink><br /><searchLink fieldCode="DE" term="%22Methanosarcina%22">Methanosarcina</searchLink><br /><searchLink fieldCode="DE" term="%22Genomes%22">Genomes</searchLink><br /><searchLink fieldCode="DE" term="%22Escherichia+coli%22">Escherichia coli</searchLink><br /><searchLink fieldCode="DE" term="%22Transcription+factors%22">Transcription factors</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Subunit D of multisubunit RNA polymerase from many species of archaea is predicted to bind one to two iron-sulfur (Fe-S) clusters, the function of which is unknown. A survey of encoded subunit D in the genomes of sequenced archaea revealed six distinct groups based on the number of complete or partial [4Fe- 4S] cluster motifs within domain 3. Only subunitDfrom strictly anaerobic archaea, including all members of the Methanosarcinales, are predicted to bind two [4Fe-4S] clusters. We report herein the purification and characterization of Methanosarcina acetivorans subunit D in complex with subunit L. Expression of subunit D and subunit L in Escherichia coli resulted in the purification of a D-L heterodimer with only partial [4Fe-4S] cluster content. Reconstitution in vitro with iron and sulfide revealed that the M. acetivorans D-L heterodimer is capable of binding two redox-active [4Fe-4S] clusters. M. acetivorans subunit D deleted of domain 3 (DΔD3) was still capable of co-purifying with subunit L but was devoid of [4Fe-4S] clusters. Affinity purification of subunit D or subunit DΔD3 from M. acetivorans resulted in the co-purification of endogenous subunit L with each tagged subunit D. Overall, these results suggest that domain 3 of subunit D is required for [4Fe-4S] cluster binding, but the [4Fe-4S] clusters and domain 3 are not required for the formation of the D-L heterodimer. However, exposure of two [4Fe-4S] cluster-containing D-L heterodimer to oxygen resulted in loss of the [4Fe-4S] clusters and subsequent protein aggregation, indicating that the [4Fe-4S] clusters influence the stability of the D-L heterodimer and therefore have the potential to regulate the assembly and/or activity of RNA polymerase in an oxidant- dependent manner. [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: 10.1074/jbc.M111.331199 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 14 StartPage: 18510 Subjects: – SubjectFull: RNA polymerases Type: general – SubjectFull: Methanosarcina Type: general – SubjectFull: Genomes Type: general – SubjectFull: Escherichia coli Type: general – SubjectFull: Transcription factors Type: general Titles: – TitleFull: Subunit D of RNA Polymerase from Methanosarcina acetivorans Contains Two Oxygen-labile [4Fe-4S] Clusters: IMPLICATIONS FOR OXIDANT-DEPENDENT REGULATION OF TRANSCRIPTION. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Lessner, Faith H. – PersonEntity: Name: NameFull: Jennings, Matthew E. – PersonEntity: Name: NameFull: Hirata, Akira – PersonEntity: Name: NameFull: Duin, Eduardus C. – PersonEntity: Name: NameFull: Lessner, Daniel J. IsPartOfRelationships: – BibEntity: Dates: – D: 25 M: 05 Text: 5/25/2012 Type: published Y: 2012 Identifiers: – Type: issn-print Value: 00219258 Numbering: – Type: volume Value: 287 – Type: issue Value: 22 Titles: – TitleFull: Journal of Biological Chemistry Type: main |
| ResultId | 1 |
