scholarly journals Metal-dependent SpoIIE oligomerization stabilizes FtsZ during asymmetric division in Bacillus subtilis

PLoS ONE ◽  
2017 ◽  
Vol 12 (3) ◽  
pp. e0174713 ◽  
Author(s):  
Ewa Król ◽  
Anabela de Sousa Borges ◽  
Malgorzata Kopacz ◽  
Dirk-Jan Scheffers
2003 ◽  
Vol 185 (13) ◽  
pp. 3905-3917 ◽  
Author(s):  
Mónica Serrano ◽  
Luísa Côrte ◽  
Jason Opdyke ◽  
Charles P. Moran, ◽  
Adriano O. Henriques

ABSTRACT During sporulation in Bacillus subtilis, the prespore-specific developmental program is initiated soon after asymmetric division of the sporangium by the compartment-specific activation of RNA polymerase sigma factor σF. σF directs transcription of spoIIIG, encoding the late forespore-specific regulator σG. Following synthesis, σG is initially kept in an inactive form, presumably because it is bound to the SpoIIAB anti-sigma factor. Activation of σG occurs only after the complete engulfment of the prespore by the mother cell. Mutations in spoIIIJ arrest sporulation soon after conclusion of the engulfment process and prevent activation of σG. Here we show that σG accumulates but is mostly inactive in a spoIIIJ mutant. We also show that expression of the spoIIIGE155K allele, encoding a form of σG that is not efficiently bound by SpoIIAB in vitro, restores σG-directed gene expression to a spoIIIJ mutant. Expression of spoIIIJ occurs during vegetative growth. However, we show that expression of spoIIIJ in the prespore is sufficient for σG activation and for sporulation. Mutations in the mother cell-specific spoIIIA locus are known to arrest sporulation just after completion of the engulfment process. Previous work has also shown that σG accumulates in an inactive form in spoIIIA mutants and that the need for spoIIIA expression for σG activation can be circumvented by the spoIIIGE155K allele. However, in contrast to the case for spoIIIJ, we show that expression of spoIIIA in the prespore does not support efficient sporulation. The results suggest that the activation of σG at the end of the engulfment process involves the action of spoIIIA from the mother cell and of spoIIIJ from the prespore.


2005 ◽  
Vol 187 (10) ◽  
pp. 3511-3520 ◽  
Author(s):  
Karen Carniol ◽  
Sigal Ben-Yehuda ◽  
Nicole King ◽  
Richard Losick

ABSTRACT SpoIIE is a dual-function protein in Bacillus subtilis that contributes to the switch from medial to polar cell division during sporulation and is responsible for activating the cell-specific transcription factor σF. SpoIIE consists of an N-terminal domain with 10 membrane-spanning segments (region I), a C-terminal phosphatase domain (region III), and a central domain (region II) of uncertain function. To investigate the role of SpoIIE in polar division, we took advantage of a system for efficiently producing polar septa during growth in a SpoIIE-dependent manner using cells engineered to produce the sporulation protein in response to an inducer. The results show that regions II and III play a critical role in polar septum formation and that specific amino acid substitutions in those regions affect the abilities of SpoIIE both to promote polar division and to localize to the division machinery. Additionally, we show that neither the phosphatase function of SpoIIE nor the N-terminal, membrane-spanning region is needed for the switch to asymmetric division.


2003 ◽  
Vol 185 (5) ◽  
pp. 1590-1598 ◽  
Author(s):  
David W. Hilbert ◽  
Patrick J. Piggot

ABSTRACT During sporulation, Bacillus subtilis undergoes an asymmetric division that results in two cells with different fates, the larger mother cell and the smaller forespore. The protein phosphatase SpoIIE, which is required for activation of the forespore-specific transcription factor σF, is also required for optimal efficiency and timing of asymmetric division. We performed a genetic screen for spoIIE mutants that were impaired in sporulation but not σF activity and isolated a strain with the mutation spoIIEV697A. The mutant exhibited a 10- to 40-fold reduction in sporulation and a sixfold reduction in asymmetric division compared to the parent. Transcription of the σF-dependent spoIIQ promoter was increased more than 10-fold and was no longer confined to the forespore. The excessive σF activity persisted even when asymmetric division was prevented. Disruption of spoIIGB did not restore asymmetric division to the spoIIEV697A mutant, indicating that the deficiency is not a consequence of predivisional activation of the mother cell-specific transcription factor σE. Deletion of the gene encoding σF (spoIIAC) restored asymmetric division; however, a mutation that dramatically reduced the number of promoters responsive to σF, spoIIAC561 (spoIIACV233 M), failed to do so. This result suggests that the block is due to expression of one of the small subset of σF-dependent genes expressed in this background or to unregulated interaction of σF with some other factor. Our results indicate that regulation of SpoIIE plays a critical role in coupling asymmetric division to σF activation in order to ensure proper spatial and temporal expression of forespore-specific genes.


2001 ◽  
Vol 183 (13) ◽  
pp. 4052-4060 ◽  
Author(s):  
Peter L. Graumann ◽  
Richard Losick

ABSTRACT Entry into sporulation in Bacillus subtilis is characterized by the formation of a polar septum, which asymmetrically divides the developing cell into forespore (the smaller cell) and mother cell compartments, and by migration of replication origin regions to extreme opposite poles of the cell. Here we show that polar septation is closely correlated with movement of replication origins to the extreme poles of the cell. Replication origin regions were visualized by the use of a cassette of tandem copies oflacO that had been inserted in the chromosome near the origin of replication and decorated with green fluorescent protein-LacI. The results showed that extreme polar placement of replication origin regions is not under sporulation control and occurred in stationary phase under conditions under which entry into sporulation was prevented. On the other hand, the formation of a polar septum, which is under sporulation control, was almost invariably associated with the presence of a replication origin region in the forespore. Moreover, cells in which the polar placement of origin regions was perturbed by deletion of the gene (smc) for the structural maintenance of chromosomes (SMC) protein were impaired in polar division. A small proportion (≈1%) of the mutant cells were able to undergo asymmetric division, but the forespore compartment of these exceptional cells was generally observed to contain a replication origin region. Immunofluorescence microscopy experiments indicated that the block in polar division caused by the absence of SMC occurred at or prior to the step of bipolar Z-ring formation by the cell division protein FtsZ. A model is discussed in which polar division is under the dual control of sporulation and an event associated with the placement of a replication origin at the cell pole.


Author(s):  
Dwight Anderson ◽  
Charlene Peterson ◽  
Gursaran Notani ◽  
Bernard Reilly

The protein product of cistron 3 of Bacillus subtilis bacteriophage Ø29 is essential for viral DNA synthesis and is covalently bound to the 5’-termini of the Ø29 DNA. When the DNA-protein complex is cleaved with a restriction endonuclease, the protein is bound to the two terminal fragments. The 28,000 dalton protein can be visualized by electron microscopy as a small dot and often is seen only when two ends are in apposition as in multimers or in glutaraldehyde-fixed aggregates. We sought to improve the visibility of these small proteins by use of antibody labeling.


Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
N Padilla-Montaño ◽  
IL Bazzocchi ◽  
L Moujir

2018 ◽  
Vol 22 (02) ◽  
pp. 82-89
Author(s):  
Friedrich von Rheinbaben ◽  
Oliver Riebe ◽  
Johanna Köhnlein ◽  
Sebastian Werner

ZusammenfassungZentrales Bauteil des Genius® 90 Therapie Systems ist der sogenannte Genius-Tank, dem die frische Dialyseflüssigkeit entnommen und in den die verbrauchte Lösung nach der Dialyse zurückgeführt wird. Daher kommt der sicheren Aufbereitung des Systems eine besondere Bedeutung zu. Hierfür wird ein Aufbereitungsverfahren unter Verwendung von UV-Licht in Kombination mit einem chemischen Desinfektionsmittel angewendet. Ziel der hier beschriebenen Untersuchung war es, die Wirkungsbreite und Wirkungstiefe dieses Aufbereitungsverfahrens unter praxisnahen Phase-3-Bedingungen zu ermitteln. Dazu wurde das Gerät mit Mikroorganismen und Viren künstlich kontaminiert und die Wirkung der einzelnen Verfahrensschritte ermittelt. Im Gegensatz zu der üblichen Vorgehensweise praxisnaher Untersuchungen machen Aufbereitungsverfahren medizinischer Geräte unter Phase-3-Kriterien meist eine neuartige Arbeitsweise erforderlich – im Falle der hier vorgestellten Untersuchung sogar die Konstruktion eines speziellen Geräts zur Platzierung von Keimträgen im Genius-Tank. Im Ergebnis konnte gezeigt werden, dass bereits UV-Licht allein sowie in Kombination mit einem chemischen Desinfektionsmittel unter praxisnahen Bedingungen eine sichere Wirksamkeit gegen Bakterien (Pseudomonas aeruginosa) und bakterielle Sporen (Bacillus subtilis), Schimmelpilze (Aspergillus brasiliensis) und Viren (Murines Parvovirus) besitzt.


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