Browsing by Subject "cytokinesis"
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Item Open Access A Wor1-Like Transcription Factor Is Essential for Virulence of Cryptococcus neoformans.(Frontiers in Cellular and Infection Microbiology, 2018-01) Paes, Hugo Costa; Derengowski, Lorena da Silveira; Peconick, Luisa Defranco Ferreira; Albuquerque, Patrícia; Pappas, Georgios Joannis; Nicola, André Moraes; Silva, Fabiana Brandão Alves; Vallim, Marcelo Afonso; Alspaugh, J Andrew; Felipe, Maria Sueli Soares; Fernandes, LarissaGti1/Pac2 transcription factors occur exclusively in fungi and their roles vary according to species, including regulating morphological transition and virulence, mating and secondary metabolism. Many of these functions are important for fungal pathogenesis. We therefore hypothesized that one of the two proteins of this family in Cryptococcus neoformans, a major pathogen of humans, would also control virulence-associated cellular processes. Elimination of this protein in C. neoformans results in reduced polysaccharide capsule expression and defective cytokinesis and growth at 37°C. The mutant loses virulence in a mouse model of cryptococcal infection and retains only partial virulence in the Galleria mellonella alternative model at 30°C. We performed RNA-Seq experiments on the mutant and found abolished transcription of genes that, in combination, are known to account for all the observed phenotypes. The protein has been named Required for cytokinesis and virulence 1 (Rcv1).Item Open Access How bacterial cell division might cheat turgor pressure - a unified mechanism of septal division in Gram-positive and Gram-negative bacteria.(BioEssays : news and reviews in molecular, cellular and developmental biology, 2017-08) Erickson, Harold PAn important question for bacterial cell division is how the invaginating septum can overcome the turgor force generated by the high osmolarity of the cytoplasm. I suggest that it may not need to. Several studies in Gram-negative bacteria have shown that the periplasm is isoosmolar with the cytoplasm. Indirect evidence suggests that this is also true for Gram-positive bacteria. In this case the invagination of the septum takes place within the uniformly high osmotic pressure environment, and does not have to fight turgor pressure. A related question is how the V-shaped constriction of Gram-negative bacteria relates to the plate-like septum of Gram-positive bacteria. I collected evidence that Gram-negative bacteria have a latent capability of forming plate-like septa, and present a model in which septal division is the basic mechanism in both Gram-positive and Gram-negative bacteria.Item Open Access The Chloroplast Tubulin Homologs FtsZA and FtsZB from the Red Alga Galdieria sulphuraria Co-assemble into Dynamic Filaments.(J Biol Chem, 2017-02-07) Chen, Yaodong; Porter, Katie; Osawa, Masaki; Augustus, Anne Marie; Milam, Sara L; Joshi, Chandra; Osteryoung, Katherine W; Erickson, Harold PFtsZ is a homolog of eukaryotic tubulin and is present in almost all bacteria and many archaea, where it is the major cytoskeletal protein in the Z ring, required for cell division. Unlike some other cell organelles of prokaryotic origin, chloroplasts have retained FtsZ as an essential component of the division machinery. However, chloroplast FtsZs have been challenging to study because they are difficult to express and purify. To this end, we have used a FATT-tag expression system to produce as soluble proteins the two chloroplast FtsZs from Galdieria sulphuraria, a thermophilic red alga. GsFtsZA and GsFtsZB assembled individually in the presence of GTP, forming large bundles of protofilaments. GsFtsZA also assembled in the presence of GDP, the first member of the FtsZ/tubulin superfamily to do so. Mixtures of GsFtsZA and GsFtsZB assembled protofilament bundles and hydrolyzed GTP at a rate approximately equal to the sum of their individual rates, suggesting a random co-assembly. GsFtsZA assembly by itself in limiting GTP gave polymers that remained stable for a prolonged time. However, when GsFtsZB was added, the co-polymers disassembled with enhanced kinetics, suggesting that the GsFtsZB regulates and enhances disassembly dynamics. GsFtsZA-mts (where mts is a membrane-targeting amphipathic helix) formed Z ring-like helices when expressed in E. coli. Co-expression of GsFtsZB (without an mts) gave co-assembly of both into similar helices. In summary, we provide biochemical evidence that GsFtsZA assembles as the primary scaffold of the chloroplast Z ring, and that GsFtsZB co-assembly enhances polymer disassembly and dynamics.Item Open Access Turgor Pressure and Possible Constriction Mechanisms in Bacterial Division.(Frontiers in microbiology, 2018-01-31) Osawa, Masaki; Erickson, Harold PBacterial cytokinesis begins with the assembly of FtsZ into a Z ring at the center of the cell. The Z-ring constriction in Gram-negative bacteria may occur in an environment where the periplasm and the cytoplasm are isoosmotic, but in Gram-positive bacteria the constriction may have to overcome a substantial turgor pressure. We address three potential sources of invagination force. (1) FtsZ itself may generate force by curved protofilaments bending the attached membrane. This is sufficient to constrict liposomes in vitro. However, this force is on the order of a few pN, and would not be enough to overcome turgor. (2) Cell wall (CW) synthesis may generate force by pushing the plasma membrane from the outside. However, this would probably require some kind of Brownian ratchet to separate the CW and membrane sufficiently to allow a glycan strand to slip in. The elastic element is not obvious. (3) Excess membrane production has the potential to contribute significantly to the invagination force. If the excess membrane is produced under the CW, it would force the membrane to bleb inward. We propose here that a combination of FtsZ pulling from the inside, and excess membrane pushing membrane inward may generate a substantial constriction force at the division site. This combined force generation mechanism may be sufficient to overcome turgor pressure. This would abolish the need for a Brownian ratchet for CW growth, and would permit CW to operate by reinforcing the constrictions generated by FtsZ and excess membrane.