Solution structure, dynamics and tetrahedral assembly of Anti-TRAP, a homo-trimeric triskelion-shaped regulator of tryptophan biosynthesis in Bacillus subtilis .

Autor: McElroy CA; Ohio State Biochemistry Program, USA.; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA., Ihms EC; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.; Biophysics Program, USA., Kumar Yadav D; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA., Holmquist ML; Ohio State Biochemistry Program, USA.; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA., Wadhwa V; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA., Wysocki VH; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.; National Resource for Native MS-Guided Structural Biology, USA., Gollnick P; Department of Biological Sciences, State University of New York, Buffalo, NY 14260, USA., Foster MP; Ohio State Biochemistry Program, USA.; Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.; Biophysics Program, USA.
Jazyk: angličtina
Zdroj: Journal of structural biology: X [J Struct Biol X] 2024 Jun 11; Vol. 10, pp. 100103. Date of Electronic Publication: 2024 Jun 11 (Print Publication: 2024).
DOI: 10.1016/j.yjsbx.2024.100103
Abstrakt: Cellular production of tryptophan is metabolically expensive and tightly regulated. The small Bacillus subtilis zinc binding Anti-TRAP protein (AT), which is the product of the yczA/rtpA gene, is upregulated in response to accumulating levels of uncharged tRNA Trp through a T-box antitermination mechanism. AT binds to the undecameric axially symmetric ring-shaped protein TRAP ( trp RNA Binding Attenuation Protein), thereby preventing it from binding to the trp leader RNA. This reverses the inhibitory effect of TRAP on transcription and translation of the trp operon. AT principally adopts two symmetric oligomeric states, a trimer (AT 3 ) featuring three-fold axial symmetry or a dodecamer (AT 12 ) comprising a tetrahedral assembly of trimers, whereas only the trimeric form binds and inhibits TRAP. We apply native mass spectrometry (nMS) and small-angle x-ray scattering (SAXS), together with analytical ultracentrifugation (AUC) to monitor the pH and concentration-dependent equilibrium between the trimeric and dodecameric structural forms of AT. In addition, we use solution nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of AT 3 , while heteronuclear 15 N relaxation measurements on both oligomeric forms of AT provide insights into the dynamic properties of binding-active AT 3 and binding-inactive AT 12 , with implications for TRAP binding and inhibition.
Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Mark P Foster reports financial support was provided by National Institutes of Health. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(© 2024 Published by Elsevier Inc.)
Databáze: MEDLINE
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