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Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis

Bacillus subtilis, as a model spore-forming Gram-positive bacterium, has been extensively used for spore germination research. Within this field, nutrient-dependent germination with specific germinant receptors (GerA, responding to L-alanine or L-valine; GerB and GerK, acting together to start spore...

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Autores principales: Grela, Anna, Jamrożek, Inga, Hubisz, Marta, Iwanicki, Adam, Hinc, Krzysztof, Kaźmierkiewicz, Rajmund, Obuchowski, Michał
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983566/
https://www.ncbi.nlm.nih.gov/pubmed/29856851
http://dx.doi.org/10.1371/journal.pone.0198561
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author Grela, Anna
Jamrożek, Inga
Hubisz, Marta
Iwanicki, Adam
Hinc, Krzysztof
Kaźmierkiewicz, Rajmund
Obuchowski, Michał
author_facet Grela, Anna
Jamrożek, Inga
Hubisz, Marta
Iwanicki, Adam
Hinc, Krzysztof
Kaźmierkiewicz, Rajmund
Obuchowski, Michał
author_sort Grela, Anna
collection PubMed
description Bacillus subtilis, as a model spore-forming Gram-positive bacterium, has been extensively used for spore germination research. Within this field, nutrient-dependent germination with specific germinant receptors (GerA, responding to L-alanine or L-valine; GerB and GerK, acting together to start spore germination process in response to AGFK) has been the most studied. There are three different variants of the GerAA subunit (299T/302S, 299A/302P, 299A/302S) of the GerA germination receptor present in B. subtilis subs. subtilis laboratory strains. According to our research, the 299A/302P one, unlike the others, interferes with the spore’s ability to germinate in L-alanine as assessed by the measurement of DPA release upon stimulation with the germinant. Multiple genetic manipulations described in this work followed by spore germination tests, together with secondary structure predictions led us to the following conclusions. First, position 302 of GerAA protein is crucial in terms of GerA germination receptor functionality; a proline residue at this position renders the GerA receptor non-functional, most probably due to a change in the protein secondary structure. Second, the 302P GerAA variant has most probably an impaired affinity to other components of GerA receptor. Together, these may explain the loss of GerA receptor’s function. Analysis of the GerAA protein should get us closer to understanding the mechanism of GerA receptor function.
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spelling pubmed-59835662018-06-16 Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis Grela, Anna Jamrożek, Inga Hubisz, Marta Iwanicki, Adam Hinc, Krzysztof Kaźmierkiewicz, Rajmund Obuchowski, Michał PLoS One Research Article Bacillus subtilis, as a model spore-forming Gram-positive bacterium, has been extensively used for spore germination research. Within this field, nutrient-dependent germination with specific germinant receptors (GerA, responding to L-alanine or L-valine; GerB and GerK, acting together to start spore germination process in response to AGFK) has been the most studied. There are three different variants of the GerAA subunit (299T/302S, 299A/302P, 299A/302S) of the GerA germination receptor present in B. subtilis subs. subtilis laboratory strains. According to our research, the 299A/302P one, unlike the others, interferes with the spore’s ability to germinate in L-alanine as assessed by the measurement of DPA release upon stimulation with the germinant. Multiple genetic manipulations described in this work followed by spore germination tests, together with secondary structure predictions led us to the following conclusions. First, position 302 of GerAA protein is crucial in terms of GerA germination receptor functionality; a proline residue at this position renders the GerA receptor non-functional, most probably due to a change in the protein secondary structure. Second, the 302P GerAA variant has most probably an impaired affinity to other components of GerA receptor. Together, these may explain the loss of GerA receptor’s function. Analysis of the GerAA protein should get us closer to understanding the mechanism of GerA receptor function. Public Library of Science 2018-06-01 /pmc/articles/PMC5983566/ /pubmed/29856851 http://dx.doi.org/10.1371/journal.pone.0198561 Text en © 2018 Grela et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Grela, Anna
Jamrożek, Inga
Hubisz, Marta
Iwanicki, Adam
Hinc, Krzysztof
Kaźmierkiewicz, Rajmund
Obuchowski, Michał
Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title_full Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title_fullStr Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title_full_unstemmed Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title_short Positions 299 and 302 of the GerAA subunit are important for function of the GerA spore germination receptor in Bacillus subtilis
title_sort positions 299 and 302 of the geraa subunit are important for function of the gera spore germination receptor in bacillus subtilis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983566/
https://www.ncbi.nlm.nih.gov/pubmed/29856851
http://dx.doi.org/10.1371/journal.pone.0198561
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