Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase

ABSTRACT Environmental awareness is an essential attribute of all organisms. The homeoviscous adaptation system of Bacillus subtilis provides a powerful experimental model for the investigation of stimulus detection and signaling mechanisms at the molecular level. These bacteria sense the order of m...

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Autores principales:	Pilar Fernández, Lucía Porrini, Daniela Albanesi, Luciano A. Abriata, Matteo Dal Peraro, Diego de Mendoza, María C. Mansilla
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	histidine kinase proline thermosensing two-component regulatory systems Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7ff4ce9ba1f34ec9b9150ce3d78db56a
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spelling	oai:doaj.org-article:7ff4ce9ba1f34ec9b9150ce3d78db56a2021-11-15T15:54:45ZTransmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase10.1128/mBio.02564-192150-7511https://doaj.org/article/7ff4ce9ba1f34ec9b9150ce3d78db56a2019-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02564-19https://doaj.org/toc/2150-7511ABSTRACT Environmental awareness is an essential attribute of all organisms. The homeoviscous adaptation system of Bacillus subtilis provides a powerful experimental model for the investigation of stimulus detection and signaling mechanisms at the molecular level. These bacteria sense the order of membrane lipids with the transmembrane (TM) protein DesK, which has an N-terminal sensor domain and an intracellular catalytic effector domain. DesK exhibits autokinase activity as well as phosphotransferase and phosphatase activities toward a cognate response regulator, DesR, that controls the expression of an enzyme that remodels membrane fluidity when the temperature drops below ∼30°C. Membrane fluidity signals are transmitted from the DesK sensor domain to the effector domain via rotational movements of a connecting 2-helix coiled coil (2-HCC). Previous molecular dynamic simulations suggested important roles for TM prolines in transducing the initial signals of membrane fluidity status to the 2-HCC. Here, we report that individual replacement of prolines in DesKs TM1 and TM5 helices by alanine (DesKPA) locked DesK in a phosphatase-ON state, abrogating membrane fluidity responses. An unbiased mutagenic screen identified the L174P replacement in the internal side of the repeated heptad of the 2-HCC structure that alleviated the signaling defects of every transmembrane DesKPA substitution. Moreover, substitutions by proline in other internal positions of the 2-HCC reestablished the kinase-ON state of the DesKPA mutants. These results imply that TM prolines are essential for finely tuned signal generation by the N-terminal sensor helices, facilitating a conformational control by the metastable 2-HCC domain of the DesK signaling state. IMPORTANCE Signal sensing and transduction is an essential biological process for cell adaptation and survival. Histidine kinases (HK) are the sensory proteins of two-component systems that control many bacterial responses to different stimuli, like environmental changes. Here, we focused on the HK DesK from Bacillus subtilis, a paradigmatic example of a transmembrane thermosensor suited to remodel membrane fluidity when the temperature drops below 30°C. DesK provides a tractable system for investigating the mechanism of transmembrane signaling, one of the majors interrogates in biology to date. Our studies demonstrate that transmembrane proline residues modulate the conformational switch of a 2-helix coiled-coil (2-HCC) structural motif that controls input-output in a variety of HK. Our results highlight the relevance of proline residues within sensor domains and could inspire investigations of their role in different signaling proteins.Pilar FernándezLucía PorriniDaniela AlbanesiLuciano A. AbriataMatteo Dal PeraroDiego de MendozaMaría C. MansillaAmerican Society for Microbiologyarticlehistidine kinaseprolinethermosensingtwo-component regulatory systemsMicrobiologyQR1-502ENmBio, Vol 10, Iss 6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	histidine kinase proline thermosensing two-component regulatory systems Microbiology QR1-502
spellingShingle	histidine kinase proline thermosensing two-component regulatory systems Microbiology QR1-502 Pilar Fernández Lucía Porrini Daniela Albanesi Luciano A. Abriata Matteo Dal Peraro Diego de Mendoza María C. Mansilla Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
description	ABSTRACT Environmental awareness is an essential attribute of all organisms. The homeoviscous adaptation system of Bacillus subtilis provides a powerful experimental model for the investigation of stimulus detection and signaling mechanisms at the molecular level. These bacteria sense the order of membrane lipids with the transmembrane (TM) protein DesK, which has an N-terminal sensor domain and an intracellular catalytic effector domain. DesK exhibits autokinase activity as well as phosphotransferase and phosphatase activities toward a cognate response regulator, DesR, that controls the expression of an enzyme that remodels membrane fluidity when the temperature drops below ∼30°C. Membrane fluidity signals are transmitted from the DesK sensor domain to the effector domain via rotational movements of a connecting 2-helix coiled coil (2-HCC). Previous molecular dynamic simulations suggested important roles for TM prolines in transducing the initial signals of membrane fluidity status to the 2-HCC. Here, we report that individual replacement of prolines in DesKs TM1 and TM5 helices by alanine (DesKPA) locked DesK in a phosphatase-ON state, abrogating membrane fluidity responses. An unbiased mutagenic screen identified the L174P replacement in the internal side of the repeated heptad of the 2-HCC structure that alleviated the signaling defects of every transmembrane DesKPA substitution. Moreover, substitutions by proline in other internal positions of the 2-HCC reestablished the kinase-ON state of the DesKPA mutants. These results imply that TM prolines are essential for finely tuned signal generation by the N-terminal sensor helices, facilitating a conformational control by the metastable 2-HCC domain of the DesK signaling state. IMPORTANCE Signal sensing and transduction is an essential biological process for cell adaptation and survival. Histidine kinases (HK) are the sensory proteins of two-component systems that control many bacterial responses to different stimuli, like environmental changes. Here, we focused on the HK DesK from Bacillus subtilis, a paradigmatic example of a transmembrane thermosensor suited to remodel membrane fluidity when the temperature drops below 30°C. DesK provides a tractable system for investigating the mechanism of transmembrane signaling, one of the majors interrogates in biology to date. Our studies demonstrate that transmembrane proline residues modulate the conformational switch of a 2-helix coiled-coil (2-HCC) structural motif that controls input-output in a variety of HK. Our results highlight the relevance of proline residues within sensor domains and could inspire investigations of their role in different signaling proteins.
format	article
author	Pilar Fernández Lucía Porrini Daniela Albanesi Luciano A. Abriata Matteo Dal Peraro Diego de Mendoza María C. Mansilla
author_facet	Pilar Fernández Lucía Porrini Daniela Albanesi Luciano A. Abriata Matteo Dal Peraro Diego de Mendoza María C. Mansilla
author_sort	Pilar Fernández
title	Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
title_short	Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
title_full	Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
title_fullStr	Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
title_full_unstemmed	Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase
title_sort	transmembrane prolines mediate signal sensing and decoding in <named-content content-type="genus-species">bacillus subtilis</named-content> desk histidine kinase
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/7ff4ce9ba1f34ec9b9150ce3d78db56a
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Transmembrane Prolines Mediate Signal Sensing and Decoding in <named-content content-type="genus-species">Bacillus subtilis</named-content> DesK Histidine Kinase

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