scholarly journals An analysis of LysM domain function in LytE when fulfilling the D,L-endopeptidase requirement for viability in Bacillus subtilis

2022 ◽  
Author(s):  
Laura Uelze
Keyword(s):  
Bacillus Subtilis ◽  
Cell Viability ◽  
Evolutionary Model ◽  
Lysm Domain ◽  
Type D ◽  
Terminal Domains

The D,L-endopeptidase requirement states that Bacillus subtilis requires either the activity of the LytE or the CwlO enzyme for viability, therefore proving that these two enzymes can complement for each other despite their very different N-terminal domains. Here, we show that another D,L-endopeptidase, LytF, can also fulfill the D,L-endopeptidase requirement for viability, when expressed from the cwlO promoter. Both LytE and LytF contain N-terminally located LysM domains, three and five respectively. However, cells expressing another very similar D,L-endopeptidase CwlS, with four LysM domains were not viable. This led us to investigate whether a LytE protein with any one of its three LysM domains permuted can fulfill the D,L-endopeptidase requirement for viability. We found that the three LysM domains are not functionally equivalent and that the N-terminally located LysM domain plays a greater role for functioning of the LytE enzyme than the subsequent domains. Based on an investigation of orthologous enzymes in 19 B. subtilis species we propose an evolutionary model describing the development of the LytE-, CwlS- and LytF-type D,L-endopeptidases and their LysM domain repeats. In summary, these results show that the LytE enzyme has been optimized to fulfill the D,L-endopeptidase requirement for cell viability of B. subtilis with regard to the number and properties of LysM domains that mediate peptidoglycan-binding.

scholarly journals A LysM Domain Intervenes in Sequential Protein-Protein and Protein-Peptidoglycan Interactions Important for Spore Coat Assembly inBacillus subtilis

2018 ◽  
Vol 201 (4) ◽  
Author(s):  
Fatima C. Pereira ◽  
Filipa Nunes ◽  
Fernando Cruz ◽  
Catarina Fernandes ◽  
Anabela L. Isidro ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Protein Interaction ◽  
Interaction Domain ◽  
Content Type ◽  
Protein Protein Interaction ◽  
Lysm Domain ◽  
Binding Function ◽  
Morphogenetic Protein ◽  
Tight Connection

ABSTRACTAt a late stage in spore development inBacillus subtilis, the mother cell directs synthesis of a layer of peptidoglycan known as the cortex between the two forespore membranes, as well as the assembly of a protective protein coat at the surface of the forespore outer membrane. SafA, the key determinant of inner coat assembly, is first recruited to the surface of the developing spore and then encases the spore under the control of the morphogenetic protein SpoVID. SafA has a LysM peptidoglycan-binding domain, SafALysM, and localizes to the cortex-coat interface in mature spores. SafALysMis followed by a region, A, required for an interaction with SpoVID and encasement. We now show that residues D10 and N30 in SafALysM, while involved in the interaction with peptidoglycan, are also required for the interaction with SpoVID and encasement. We further show that single alanine substitutions on residues S11, L12, and I39 of SafALysMthat strongly impair binding to purified cortex peptidoglycan affect a later stage in the localization of SafA that is also dependent on the activity of SpoVE, a transglycosylase required for cortex formation. The assembly of SafA thus involves sequential protein-protein and protein-peptidoglycan interactions, mediated by the LysM domain, which are required first for encasement then for the final localization of the protein in mature spores.IMPORTANCEBacillus subtilisspores are encased in a multiprotein coat that surrounds an underlying peptidoglycan layer, the cortex. How the connection between the two layers is enforced is not well established. Here, we elucidate the role of the peptidoglycan-binding LysM domain, present in two proteins, SafA and SpoVID, that govern the localization of additional proteins to the coat. We found that SafALysMis a protein-protein interaction module during the early stages of coat assembly and a cortex-binding module at late stages in morphogenesis, with the cortex-binding function promoting a tight connection between the cortex and the coat. In contrast, SpoVIDLysMfunctions only as a protein-protein interaction domain that targets SpoVID to the spore surface at the onset of coat assembly.

scholarly journals Effects of Bacillus Subtilis-Fermented White Sword Bean Extract on Adipogenesis and Lipolysis of 3T3-L1 Adipocytes

Foods ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1423
Author(s):  
Yujeong Choi ◽  
Da-Som Kim ◽  
Min-Chul Lee ◽  
Seulgi Park ◽  
Joo-Won Lee ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Cell Viability ◽  
Dietary Supplement ◽  
Free Glycerol ◽  
Glycerol Content ◽  
Treatment Groups ◽  
Protein Levels ◽  
Triglyceride Accumulation ◽  
Before And After

To investigate the adipogenesis and lipolysis effects of the Bacillus subtilis-fermented white sword bean extract (FWSBE) on 3T3-L1 adipocytes, we treated 3T3-L1 preadipocytes before and after differentiation with FWSBE and measured triglyceride, free glycerol, mRNA, and protein levels. First, FWSBE reduced the cell viability of 3T3-L1 pre-adipocytes under 1000 µg/mL conditions. Triglyceride accumulation in 3T3-L1 pre-adipocytes was suppressed, and free glycerol content in mature 3T3-L1 adipocytes was increased in the FWSBE treatment groups, indicating that FWSBE has anti-obesity effects. Further, FWSBE suppressed adipogenesis in 3T3-L1 pre-adipocytes by lowering the protein levels of C/EBPα, PPARγ, and FAS and increasing the level of pACC and pAMPK. Additionally, FWSBE promoted lipolysis in mature 3T3-L1 adipocytes by increasing the transcription levels of Ppara, Acox, and Lcad and the protein levels of pHSL and ATGL. Thus, we suggest that FWSBE can be a potential dietary supplement because of its anti-obesity properties.

scholarly journals Cytochemical studies of cell viability and gene expression in Bacillus subtilis macrofibres

Microbiology ◽  
1997 ◽  
Vol 143 (12) ◽  
pp. 3713-3721 ◽  
Author(s):  
N. H. Mendelson ◽  
B. Salhi ◽  
K. E. Sundahl
Keyword(s):  
Gene Expression ◽  
Bacillus Subtilis ◽  
Cell Viability

scholarly journals The PrpC Serine-Threonine Phosphatase and PrkC Kinase Have Opposing Physiological Roles in Stationary-Phase Bacillus subtilis Cells

2002 ◽  
Vol 184 (22) ◽  
pp. 6109-6114 ◽  
Author(s):  
Tatiana A. Gaidenko ◽  
Tae-Jong Kim ◽  
Chester W. Price
Keyword(s):  
Bacillus Subtilis ◽  
Stationary Phase ◽  
Cell Viability ◽  
Cell Density ◽  
Elongation Factor ◽  
Pleiotropic Effects ◽  
General Stress ◽  
Factor G

ABSTRACT Loss of the PrpC serine-threonine phosphatase and the associated PrkC kinase of Bacillus subtilis were shown to have opposite effects on stationary-phase physiology by differentially affecting cell density, cell viability, and accumulation of β-galactosidase from a general stress reporter fusion. These pleiotropic effects suggest that PrpC and PrkC have important regulatory roles in stationary-phase cells. Elongation factor G (EF-G) was identified as one possible target of the PrpC and PrkC pair in vivo, and purified PrpC and PrkC manifested the predicted phosphatase and kinase activities against EF-G in vitro.

scholarly journals Interaction between Coat Morphogenetic Proteins SafA and SpoVID

2006 ◽  
Vol 188 (22) ◽  
pp. 7731-7741 ◽  
Author(s):  
Teresa Costa ◽  
Anabela L. Isidro ◽  
Charles P. Moran ◽  
Adriano O. Henriques
Keyword(s):  
Bacillus Subtilis ◽  
Amino Acid ◽  
Important Determinant ◽  
Structural Components ◽  
Lysm Domain ◽  
Amino Acid Region ◽  
Acid Region

ABSTRACT Morphogenetic proteins such as SpoVID and SafA govern assembly of the Bacillus subtilis endospore coat by guiding the various protein structural components to the surface of the developing spore. Previously, a screen for peptides able to interact with SpoVID led to the identification of a PYYH motif present in the C-terminal half of the SafA protein and to the subsequent demonstration that SpoVID and SafA directly interact. spoVID and safA spores show deficiencies in coat assembly and are lysozyme susceptible. Both proteins, orthologs of which are found in all Bacillus species, have LysM domains for peptidoglycan binding and localize to the cortex-coat interface. Here, we show that the interaction between SafA and SpoVID involves the PYYH motif (region B) but also a 13-amino-acid region (region A) just downstream of the N-terminal LysM domain of SafA. We show that deletion of region B does not block the interaction of SafA with SpoVID, nor does it bring about spore susceptibility to lysozyme. Nevertheless, it appears to reduce the interaction and affects the complex. In contrast, lesions in region A impaired the interaction of SafA with SpoVID in vitro and, while not affecting the accumulation of SafA in vivo, interfered with the localization of SafA around the developing spore, causing aberrant assembly of the coat and lysozyme sensitivity. A peptide corresponding to region A interacts with SpoVID, suggesting that residues within this region directly contact SpoVID. Since region A is highly conserved among SafA orthologs, this motif may be an important determinant of coat assembly in the group of Bacillus spore formers.

scholarly journals Wall Teichoic Acid Polymers Are Dispensable for Cell Viability in Bacillus subtilis

2006 ◽  
Vol 188 (23) ◽  
pp. 8313-8316 ◽  
Author(s):  
Michael A. D'Elia ◽  
Kathryn E. Millar ◽  
Terry J. Beveridge ◽  
Eric D. Brown
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Bacillus Subtilis ◽  
Cell Viability ◽  
Teichoic Acid ◽  
Acid Synthesis ◽  
Extensive Literature ◽  
Wall Teichoic Acid ◽  
First Time ◽  
Similar Gene

ABSTRACT An extensive literature has established that the synthesis of wall teichoic acid in Bacillus subtilis is essential for cell viability. Paradoxically, we have recently shown that wall teichoic acid biogenesis is dispensable in Staphylococcus aureus (M. A. D'Elia, M. P. Pereira, Y. S. Chung, W. Zhao, A. Chau, T. J. Kenney, M. C. Sulavik, T. A. Black, and E. D. Brown, J. Bacteriol. 188:4183-4189, 2006). A complex pattern of teichoic acid gene dispensability was seen in S. aureus where the first gene (tarO) was dispensable and later acting genes showed an indispensable phenotype. Here we show, for the first time, that wall teichoic acid synthesis is also dispensable in B. subtilis and that a similar gene dispensability pattern is seen where later acting enzymes display an essential phenotype, while the gene tagO, whose product catalyzes the first step in the pathway, could be deleted to yield viable mutants devoid of teichoic acid in the cell wall.

scholarly journals The Crystal Structure of Bacillus subtilis YycI Reveals a Common Fold for Two Members of an Unusual Class of Sensor Histidine Kinase Regulatory Proteins

2007 ◽  
Vol 189 (8) ◽  
pp. 3290-3295 ◽  
Author(s):  
Eugenio Santelli ◽  
Robert C. Liddington ◽  
Michael A. Mohan ◽  
James A. Hoch ◽  
Hendrik Szurmant
Keyword(s):  
Crystal Structure ◽  
Bacillus Subtilis ◽  
Sequence Homology ◽  
Histidine Kinase ◽  
Direct Interaction ◽  
Regulatory Proteins ◽  
Sensor Histidine Kinase ◽  
Periplasmic Proteins ◽  
Terminal Domains

ABSTRACT YycI and YycH are two membrane-anchored periplasmic proteins that regulate the essential Bacillus subtilis YycG histidine kinase through direct interaction. Here we present the crystal structure of YycI at a 2.9-Å resolution. YycI forms a dimer, and remarkably the structure resembles that of the two C-terminal domains of YycH despite nearly undetectable sequence homology (10%) between the two proteins.

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