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

Anna Grela , Inga Jamrożek , Marta Hubisz , Adam Iwanicki , Krzysztof Hinc , Rajmund Kaźmierkiewicz , Michał Obuchowski

Abstract

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.
Author Anna Grela (IFB)
Anna Grela,,
- Intercollegiate Faculty of Biotechnology UG
, Inga Jamrożek (IFB)
Inga Jamrożek,,
- Intercollegiate Faculty of Biotechnology UG
, Marta Hubisz (IFB)
Marta Hubisz,,
- Intercollegiate Faculty of Biotechnology UG
, Adam Iwanicki (IFB / DMedB)
Adam Iwanicki,,
- Department of Medical Biotechnology
, Krzysztof Hinc (IFB / DMedB)
Krzysztof Hinc,,
- Department of Medical Biotechnology
, Rajmund Kaźmierkiewicz (IFB / IB / LBSS)
Rajmund Kaźmierkiewicz,,
- Laboratory of Biomolecular Systems Simulations
, Michał Obuchowski (IFB / DMedB)
Michał Obuchowski,,
- Department of Medical Biotechnology
Journal seriesPlos One, ISSN 1932-6203, (A 35 pkt)
Issue year2018
Vol13
No6
Pages1-15
Publication size in sheets0.7
DOIDOI:10.1371/journal.pone.0198561
URL https://doi.org/10.1371/journal.pone.0198561
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)40
ScoreMinisterial score = 35.0, 09-10-2018, ArticleFromJournal
Ministerial score (2013-2016) = 40.0, 09-10-2018, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 2.806 (2) - 2016=3.394 (5)
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