Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein

The stringent response, regulated by the bifunctional (p)ppGpp synthetase/hydrolase Rel in mycobacteria, is critical for long-term survival of the drug-tolerant dormant state of Mycobacterium tuberculosis. During amino acid starvation, MtRel senses a drop in amino acid concentration and synthesizes...

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Main Authors: Shin, Joon, Singal, Bharti, Manimekalai, Malathy Sony Subramanian, Wei Chen, Ming, Ragunathan, Priya, Grüber, Gerhard
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2021
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Online Access:https://hdl.handle.net/10356/154271
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-1542712021-12-31T13:25:24Z Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein Shin, Joon Singal, Bharti Manimekalai, Malathy Sony Subramanian Wei Chen, Ming Ragunathan, Priya Grüber, Gerhard School of Biological Sciences Science::Biological sciences ACT Domain Mycobacterium tuberculosis The stringent response, regulated by the bifunctional (p)ppGpp synthetase/hydrolase Rel in mycobacteria, is critical for long-term survival of the drug-tolerant dormant state of Mycobacterium tuberculosis. During amino acid starvation, MtRel senses a drop in amino acid concentration and synthesizes the messengers pppGpp and ppGpp, collectively called (p)ppGpp. Here, we investigate the role of the regulatory 'Aspartokinase, Chorismate mutase and TyrA' (ACT) domain in MtRel. Using NMR spectroscopy approaches, we report the high-resolution structure of dimeric MtRel ACT which selectively binds to valine out of all other branched-chain amino acids tested. A set of MtRel ACT mutants were generated to identify the residues required for maintaining the head-to-tail dimer. Through NMR titrations, we determined the crucial residues for binding of valine and show structural rearrangement of the MtRel ACT dimer in the presence of valine. This study suggests the direct involvement of amino acids in (p)ppGpp accumulation mediated by MtRel independent to interactions with stalled ribosomes. Database Structural data are available in the PDB database under the accession number 6LXG. Ministry of Education (MOE) We would like to thank the Singapore Ministry ofEducation Academic Research Fund Tier 1 (Rg137/15)for the funding to GG. BS thanks the Nanyang Tech-nological University, Singapore, for awarding her aPhD Scholarship during her studies. 2021-12-16T07:34:47Z 2021-12-16T07:34:47Z 2021 Journal Article Shin, J., Singal, B., Manimekalai, M. S. S., Wei Chen, M., Ragunathan, P. & Grüber, G. (2021). Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein. FEBS Journal, 288, 2377-2397. https://dx.doi.org/10.1111/febs.15600 1742-464X https://hdl.handle.net/10356/154271 10.1111/febs.15600 33067840 2-s2.0-85096745401 288 2377 2397 en Rg137/15 FEBS Journal © 2020 Federation of European Biochemical Societies. All rights reserved.
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Science::Biological sciences
ACT Domain
Mycobacterium tuberculosis
spellingShingle Science::Biological sciences
ACT Domain
Mycobacterium tuberculosis
Shin, Joon
Singal, Bharti
Manimekalai, Malathy Sony Subramanian
Wei Chen, Ming
Ragunathan, Priya
Grüber, Gerhard
Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
description The stringent response, regulated by the bifunctional (p)ppGpp synthetase/hydrolase Rel in mycobacteria, is critical for long-term survival of the drug-tolerant dormant state of Mycobacterium tuberculosis. During amino acid starvation, MtRel senses a drop in amino acid concentration and synthesizes the messengers pppGpp and ppGpp, collectively called (p)ppGpp. Here, we investigate the role of the regulatory 'Aspartokinase, Chorismate mutase and TyrA' (ACT) domain in MtRel. Using NMR spectroscopy approaches, we report the high-resolution structure of dimeric MtRel ACT which selectively binds to valine out of all other branched-chain amino acids tested. A set of MtRel ACT mutants were generated to identify the residues required for maintaining the head-to-tail dimer. Through NMR titrations, we determined the crucial residues for binding of valine and show structural rearrangement of the MtRel ACT dimer in the presence of valine. This study suggests the direct involvement of amino acids in (p)ppGpp accumulation mediated by MtRel independent to interactions with stalled ribosomes. Database Structural data are available in the PDB database under the accession number 6LXG.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Shin, Joon
Singal, Bharti
Manimekalai, Malathy Sony Subramanian
Wei Chen, Ming
Ragunathan, Priya
Grüber, Gerhard
format Article
author Shin, Joon
Singal, Bharti
Manimekalai, Malathy Sony Subramanian
Wei Chen, Ming
Ragunathan, Priya
Grüber, Gerhard
author_sort Shin, Joon
title Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
title_short Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
title_full Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
title_fullStr Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
title_full_unstemmed Atomic structure of, and valine binding to the regulatory ACT domain of the Mycobacterium tuberculosis Rel protein
title_sort atomic structure of, and valine binding to the regulatory act domain of the mycobacterium tuberculosis rel protein
publishDate 2021
url https://hdl.handle.net/10356/154271
_version_ 1722355346924634112