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The Rel stringent factor from Thermus thermophilus: crystallization and X-ray analysis
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS).
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS).
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2019 (English)In: Acta Crystallographica Section F : Structural Biology Communications, ISSN 2053-230X, Vol. 75, p. 561-569Article in journal (Refereed) Published
Abstract [en]

The stringent response, controlled by (p)ppGpp, enables bacteria to trigger a strong phenotypic resetting that is crucial to cope with adverse environmental changes and is required for stress survival and virulence. In the bacterial cell, (p)ppGpp levels are regulated by the concerted opposing activities of RSH (RelA/SpoT homologue) enzymes that can transfer a pyrophosphate group of ATP to the 3′ position of GDP (or GTP) or remove the 3′ pyrophosphate moiety from (p)ppGpp. Bifunctional Rel enzymes are notoriously difficult to crystallize owing to poor stability and a propensity for aggregation, usually leading to a loss of biological activity after purification. Here, the production, biochemical analysis and crystallization of the bifunctional catalytic region of the Rel stringent factor from Thermus thermophilus (RelTtNTD) in the resting state and bound to nucleotides are described. RelTt and RelTtNTD are monomers in solution that are stabilized by the binding of Mn2+ and mellitic acid. RelTtNTD crystallizes in space group P4122, with unit-cell parameters a = b = 88.4, c = 182.7 Å, at 4°C and in space group P41212, with unit-cell parameters a = b = 105.7, c = 241.4 Å, at 20°C.

Place, publisher, year, edition, pages
International Union of Crystallography , 2019. Vol. 75, p. 561-569
Keywords [en]
stringent response, Rel/RelA/SpoT, (p)ppGpp, bacterial alarmone, Thermus thermophilus
National Category
Microbiology in the medical area
Identifiers
URN: urn:nbn:se:umu:diva-162870DOI: 10.1107/S2053230X19010628ISI: 000480337300007PubMedID: 31397328OAI: oai:DiVA.org:umu-162870DiVA, id: diva2:1348869
Available from: 2019-09-05 Created: 2019-09-05 Last updated: 2019-09-05Bibliographically approved

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Takada, HirakuHauryliuk, Vasili

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Department of Molecular Biology (Faculty of Medicine)Molecular Infection Medicine Sweden (MIMS)
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