scholarly journals Localized Cardiolipin Synthesis is Required for the Assembly of MreB During the Polarized Cell Division of Chlamydia trachomatis

2022 ◽  
Author(s):  
Scot P Ouellette ◽  
Laura A Fisher-Marvin ◽  
McKenna Harpring ◽  
Junghoon Lee ◽  
Elizabeth A Rucks ◽  
...  

Pathogenic Chlamydia species are coccoid bacteria that use the rod-shape determining protein MreB to direct septal peptidoglycan synthesis during their polarized cell division process. How the site of polarized budding is determined in this bacterium, where contextual features like membrane curvature are seemingly identical, is unclear. We hypothesized that the accumulation of the phospholipid, cardiolipin (CL), in specific regions of the cell membrane induces localized membrane changes that trigger the recruitment of MreB to the site where the bud will arise. To test this, we ectopically expressed cardiolipin synthase (Cls) and observed a polar distribution for this enzyme in Chlamydia trachomatis. In early division intermediates, Cls was restricted to the bud site where MreB is localized and peptidoglycan synthesis is initiated. The localization profile of Cls throughout division mimicked the distribution of lipids that stain with NAO, a dye that labels CL. Treatment of Chlamydia with 3-,6-dinonylneamine (diNN), an antibiotic targeting CL-containing membrane domains, resulted in redistribution of Cls and NAO-staining phospholipids. In addition, MreB localization was altered by diNN treatment, suggesting an upstream regulatory role for CL-containing membranes in directing the assembly of MreB. This hypothesis is consistent with the observation that the clustered localization of Cls is not dependent upon MreB function or peptidoglycan synthesis. Furthermore, expression of a CL-binding protein at the inner membrane of C. trachomatis dramatically inhibited bacterial growth supporting the importance of CL in the division process. Our findings implicate a critical role for localized CL synthesis in driving MreB assembly at the bud site during the polarized cell division of Chlamydia.

2000 ◽  
Vol 68 (2) ◽  
pp. 708-715 ◽  
Author(s):  
W. J. Brown ◽  
D. D. Rockey

ABSTRACT The process of chlamydial cell division has not been thoroughly investigated. The lack of detectable peptidoglycan and the absence of an FtsZ homolog within chlamydiae suggest an unusual mechanism for the division process. Our laboratory has identified an antigen (SEP antigen) localized to a ring-like structure at the apparent septum within dividing chlamydial reticulate bodies (RB). Antisera directed against SEP show similar patterns of antigen distribution inChlamydia trachomatis and Chlamydia psittaciRB. In contrast to localization in RB, SEP in elementary bodies appears diffuse and irregular, suggesting that the distribution of the antigen is developmental-stage specific. Treatment of chlamydiae with inhibitors of peptidoglycan synthesis or culture of chlamydiae in medium lacking tryptophan leads to the formation of nondividing, aberrant RB. Staining of aberrant RB with anti-SEP reveals a marked redistribution of the antigen. Within C. trachomatis-infected cells, ampicillin treatment leads to high levels of SEP accumulation at the periphery of aberrant RB, while inC. psittaci, treatment causes SEP to localize to distinct punctate sites within the bacteria. Aberrancy produced via tryptophan depletion results in a different pattern of SEP distribution. In either case, the reversal of aberrant formation results in the production of normal RB and a redistribution of SEP to the apparent plane of bacterial division. Collectively these studies identify a unique chlamydial-genus-common and developmental-stage-specific antigen that may be associated with RB division.


2021 ◽  
Vol 22 (15) ◽  
pp. 8350
Author(s):  
Naďa Labajová ◽  
Natalia Baranova ◽  
Miroslav Jurásek ◽  
Robert Vácha ◽  
Martin Loose ◽  
...  

DivIVA is a protein initially identified as a spatial regulator of cell division in the model organism Bacillus subtilis, but its homologues are present in many other Gram-positive bacteria, including Clostridia species. Besides its role as topological regulator of the Min system during bacterial cell division, DivIVA is involved in chromosome segregation during sporulation, genetic competence, and cell wall synthesis. DivIVA localizes to regions of high membrane curvature, such as the cell poles and cell division site, where it recruits distinct binding partners. Previously, it was suggested that negative curvature sensing is the main mechanism by which DivIVA binds to these specific regions. Here, we show that Clostridioides difficile DivIVA binds preferably to membranes containing negatively charged phospholipids, especially cardiolipin. Strikingly, we observed that upon binding, DivIVA modifies the lipid distribution and induces changes to lipid bilayers containing cardiolipin. Our observations indicate that DivIVA might play a more complex and so far unknown active role during the formation of the cell division septal membrane.


2001 ◽  
Vol 114 (1) ◽  
pp. 207-218 ◽  
Author(s):  
C.G. Padmashree ◽  
U. Surana

In the budding yeast Saccharomyces cerevisiae, a prospective mother normally commences the formation of a daughter (the bud) only in the G(1) phase of the cell division cycle. This suggests a strict temporal regulation of the processes that initiate the formation of a new bud. Using cortical localization of bud site components Spa2 and Bni1 as an indicator of bud site assembly, we show that cells assemble a bud site following inactivation of the Cdc28-Clb mitotic kinase but prior to START. Interestingly, an untimely inactivation of the mitotic kinase is sufficient to drive cells to assemble a new bud site inappropriately in G(2) or M phases. The induction of Cdc28/Clb kinase activity in G(1), on the other hand, dramatically reduces a cell's ability to construct an incipient bud site. Our findings strongly suggest that the Cdc28-Clb kinase plays a critical role in the mechanism that restricts the timing of bud formation to the G(1) phase of the cell cycle.


Biomolecules ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1463
Author(s):  
Xochitl Morales-de la Cruz ◽  
Alejandra Mandujano-Chávez ◽  
Daniel R. Browne ◽  
Timothy P. Devarenne ◽  
Lino Sánchez-Segura ◽  
...  

Botryococcus braunii produce liquid hydrocarbons able to be processed into combustion engine fuels. Depending on the growing conditions, the cell doubling time can be up to 6 days or more, which is a slow growth rate in comparison with other microalgae. Few studies have analyzed the cell cycle of B. braunii. We did a bioinformatic comparison between the protein sequences for retinoblastoma and cyclin-dependent kinases from the A (Yamanaka) and B (Showa) races, with those sequences from other algae and Arabidopsis thaliana. Differences in the number of cyclin-dependent kinases and potential retinoblastoma phosphorylation sites between the A and B races were found. Some cyclin-dependent kinases from both races seemed to be phylogenetically more similar to A. thaliana than to other microalgae. Microscopic observations were done using several staining procedures. Race A colonies, but not race B, showed some multinucleated cells without chlorophyll. An active mitochondrial net was detected in those multinucleated cells, as well as being defined in polyphosphate bodies. These observations suggest differences in the cell division processes between the A and B races of B. braunii.


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