Influence of cell-wall thickness on cell division: electron microscopic study with Bacillus cereus

1973 ◽  
Vol 19 (2) ◽  
pp. 217-221 ◽  
Author(s):  
K. L. Chung

Bacillus cereus that had been exposed to chloramphenicol for 0 and 90 min synthesized walls whose thickness increased with time. These cells were washed and resuspended in fresh growth medium. Cell division was examined by electron microscopy. In untreated cells, a slight invagination of the cytoplasmic membrane with deposit of cross-wall material marked the onset of cell division. The cross wall grew inward until the septum was about [Formula: see text] completed. This was followed by splitting and bifurcation of the outer edge of the cross wall, which in turn separated the peripheral wall scars. Cross-wall penetration with further splitting continued until the daughter cells were completely partitioned. The peripheral wall scars that were located at the junction of peripheral and end walls at the time of cell separation indicated that the zone of cell division was probably not a region of active cell elongation. In cells treated with chloramphenicol for 90 min, cross-wall initiation and completion all occurred beneath a thickened peripheral wall. The circumferential portion of the peripheral wall at the zone of cell division appeared to be responsible for the hindrance of cell separation. This thick layer subsequently severed at three or four sites to allow the separation of daughter cells. For cells that had initiated cross-wall invagination at the time when chloramphenicol was added, accumulation of a large amount of cross-wall material at the zone of cell division was observed. Upon removal of the antibiotic, cells abandoned the old and initiated a new dividing site. For cells that had already completed cross-wall formation at the time of chloramphenicol treatment, the increase in wall thickness did not appear to interfere with cell separation. Models for the different stages of cell division in all these bacteria were presented.

2004 ◽  
Vol 15 (8) ◽  
pp. 3903-3914 ◽  
Author(s):  
Nick Dekker ◽  
Dave Speijer ◽  
Christian H. Grün ◽  
Marlene van den Berg ◽  
Annett de Haan ◽  
...  

Cell division in the fission yeast Schizosaccharomyces pombe yields two equal-sized daughter cells. Medial fission is achieved by deposition of a primary septum flanked by two secondary septa within the dividing cell. During the final step of cell division, cell separation, the primary septum is hydrolyzed by an endo-(1,3)-β-glucanase, Eng1p. We reasoned that the cell wall material surrounding the septum, referred to here as the septum edging, also must be hydrolyzed before full separation of the daughter cells can occur. Because the septum edging contains (1,3)-α-glucan, we investigated the cellular functions of the putative (1,3)-α-glucanases Agn1p and Agn2p. Whereas agn2 deletion results in a defect in endolysis of the ascus wall, deletion of agn1 leads to clumped cells that remained attached to each other by septum-edging material. Purified Agn1p hydrolyzes (1,3)-α-glucan predominantly into pentasaccharides, indicating an endo-catalytic mode of hydrolysis. Furthermore, we show that the transcription factors Sep1p and Ace2p regulate both eng1 and agn1 expression in a cell cycle-dependent manner. We propose that Agn1p acts in concert with Eng1p to achieve efficient cell separation, thereby exposing the secondary septa as the new ends of the daughter cells.


1970 ◽  
Vol 16 (11) ◽  
pp. 1027-1031 ◽  
Author(s):  
S. F. Kennedy ◽  
R. R. Colwell ◽  
G. B. Chapman

The structure of Vibrio marinus strain PS-207 was studied by both phase and electron microscopy. It was found to possess a trilaminar plasma membrane and cell wall. Membrane-bounded subunits containing DNA-like material were found dispersed throughout the cytoplasm. Giant round forms or "macrospheres" were observed in all growth stages. The size, shape, and construction of the "macrospheres" showed some variation, but could not be related to culture age. Studies of cell division in V. marinus strain PS-207 indicate the primary mechanism to be a synthesis and centripetal deposition of plasma membrane with a concomitant or subsequent synthesis and centripetal deposition of cross wall material.


1967 ◽  
Vol 13 (4) ◽  
pp. 341-350 ◽  
Author(s):  
K. L. Chung

The pattern of cell wall synthesis as measured by the incorporation of tritiated alanine into the cell wall of Bacillus cereus, and the number of synthesizing sites in the cell wall were studied by the direct and the reverse autoradiographic labelling methods.In the absence of chloramphenicol, the new cell wall was initiated at two or three segments, and later increased to four or five segments which continued to elongate but not to increase in number until the bacilli had made preparation for cell division. Shortly before the centripetal growth of the cell wall and constriction to separate daughter cells, two to three more new wall-segments were added to those already present. The second and third generation cells retained some old wall-segments from the first-generation mother, which remained as discrete clusters of grains, and could easily be distinguished from the new segments.In the presence of chloramphenicol, the new wall was initiated at 8 to 10 sites. Further incubation resulted in the uniform incorporation of labels at multiple sites along the entire cell length.The patterns of new wall replication as studied by the two methods were compared. To account for the difference in synthesizing sites when chloramphenicol is present, it is suggested that the cells have either used the maximum number of sites or have completely bypassed all the sites and allowed the tritiated alanine to diffuse into the wall to become incorporated.


1993 ◽  
Vol 71 (8) ◽  
pp. 1032-1038 ◽  
Author(s):  
A. Tsuneda ◽  
S. Murakami ◽  
Lynne Sigler ◽  
Y. Hiratsuka

An arthroconidial anamorphic fungus occurred in pupal chambers of the mountain pine beetle (Dendroctonus ponderosae) in lodgepole pine. No teleomorph was found and no suitable form genus was available for its disposition. However, in cultural characteristics it closely resembled the group 1 arthroconidial fungi, defined by other researchers as probable basidiomycete anamorphs. Septa of the pupal-chamber fungus and several of the group 1 isolates were of the dolipore–parenthesome type. Conidial separation was by septum schizolysis. Cell separation was initiated by disintegration of the dolipore wall, followed by progressive shrinkage of the fused dolipore wall. The parenthesomes retracted towards the dolipore wall and the triangular areas of the cross wall dissolved. The cross wall then split centripetally along the median electron-light layer to complete cell separation. The pupal-chamber fungus was also compared with Phlebia radiata (group 2) and with groups 3 and 4 strains of other researchers. Mauginiella scaettae was confirmed to have simple septa; thus this genus fails to accommodate arthroconidial basidiomycete anamorphs. Key words: basidiomycete anamorphs, dolipore–parenthesome septa, septum schizolysis, arthroconidiogenesis, Phlebia radiata, Mauginiella scaettae.


1971 ◽  
Vol 17 (12) ◽  
pp. 1561-1565 ◽  
Author(s):  
K. L. Chung

Bacillus cereus incubated for 4 h in a synthetic medium containing chloramphenicol was observed to form cell walls 2 to 3 times as thick as those from control cells growing in the same medium containing no antibiotic. Then the cells were washed and reincubated in fresh synthetic medium and the ultra-structural changes in the thickened walls during cell growth and elongation were examined by electron microscopy. After incubation for 20 min, multiple ruptured sites and internal fractures appeared randomly on the surface of the thickened cell wall. Large and small pieces of thickened wall fragments soon "peeled off" from the surface, leaving behind a deeper layer of wall material. Normal cell growth and elongation resumed after partial removal of the thickened cell wall. After several generations, thickened wall fragments were not observed on the surface of daughter cells.


1998 ◽  
Vol 62 (4) ◽  
pp. 1371-1414 ◽  
Author(s):  
Peter Giesbrecht ◽  
Thomas Kersten ◽  
Heinrich Maidhof ◽  
Jörg Wecke

SUMMARY The primary goal of this review is to provide a compilation of the complex architectural features of staphylococcal cell walls and of some of their unusual morphogenetic traits including the utilization of murosomes and two different mechanisms of cell separation. Knowledge of these electron microscopic findings may serve as a prerequisite for a better understanding of the sophisticated events which lead to penicillin-induced death. For more than 50 years there have been controversial disputes about the mechanisms by which penicillin kills bacteria. Many hypotheses have tried to explain this fatal event biochemically and mainly via bacteriolysis. However, indications that penicillin-induced death of staphylococci results from overall biochemical defects or from a fatal attack of bacterial cell walls by bacteriolytic murein hydrolases were not been found. Rather, penicillin, claimed to trigger the activity of murein hydrolases, impaired autolytic wall enzymes of staphylococci. Electron microscopic investigations have meanwhile shown that penicillin-mediated induction of seemingly minute cross wall mistakes is the very reason for this killing. Such “morphogenetic death” taking place at predictable cross wall sites and at a predictable time is based on the initiation of normal cell separations in those staphylococci in which the completion of cross walls had been prevented by local penicillin-mediated impairment of the distribution of newly synthesized peptidoglycan; this death occurs because the high internal pressure of the protoplast abruptly kills such cells via ejection of some cytoplasm during attempted cell separation. An analogous fatal onset of cell partition is considered to take place without involvement of a detectable quantity of autolytic wall enzymes (“mechanical cell separation”). The most prominent feature of penicillin, the disintegration of bacterial cells via bacteriolysis, is shown to represent only a postmortem process resulting from shrinkage of dead cells and perturbation of the cytoplasmic membrane. Several schematic drawings have been included in this review to facilitate an understanding of the complex morphogenetic events.


2005 ◽  
Vol 16 (4) ◽  
pp. 2003-2017 ◽  
Author(s):  
Maria Luisa Alonso-Nuñez ◽  
Hanbing An ◽  
Ana Belén Martín-Cuadrado ◽  
Sapna Mehta ◽  
Claudia Petit ◽  
...  

Schizosaccharomyces pombe cells divide by medial fission through contraction of an actomyosin ring and deposition of a multilayered division septum that must be cleaved to release the two daughter cells. Here we describe the identification of seven genes (adg1+, adg2+, adg3+, cfh4+, agn1+, eng1+, and mid2+) whose expression is induced by the transcription factor Ace2p. The expression of all of these genes varied during the cell cycle, maximum transcription being observed during septation. At least three of these proteins (Eng1p, Agn1p, and Cfh4p) localize to a ring-like structure that surrounds the septum region during cell separation. Deletion of the previously uncharacterized genes was not lethal to the cells, but produced defects or delays in cell separation to different extents. Electron microscopic observation of mutant cells indicated that the most severe defect is found in eng1Δ agn1Δ cells, lacking the Eng1p endo-β-1,3-glucanase and the Agn1p endo-α-glucanase. The phenotype of this mutant closely resembled that of ace2Δ mutants, forming branched chains of cells. This suggests that these two proteins are the main activities required for cell separation to be completed.


1964 ◽  
Vol 10 (3) ◽  
pp. 473-482 ◽  
Author(s):  
K. L. Chung ◽  
R. Z. Hawirko ◽  
P. K. Isaac

Cell wall replication in E. coli and S. faecalis was studied by differential labelling of living cells with fluorescent and non-fluorescent antibody.In E. coli the initial step in cell division was the formation of a cross wall at the cell equator, followed by the appearance of new cell wall on either side of the cross wall. The process was repeated in sequence at subsequent sites in the polar, the subcentral, and the subpolar areas. Constriction occurred at random so that the divided parent cells were composed of several daughter cells.A polar type of unidirectional cell wall growth and elongation was also observed in E. coli. It was initiated by the synthesis of a ring of new cell wall material around the polar tip. A second ring was then formed at the subpolar area during the rapid enlargement of the first ring in a single direction.Evidence shows that cell wall synthesis is independent of cell division and that in E. coli, it is initiated at multiple but specific sites within the cell and not by diffuse intercalation of old and new walls.Contrary to the synthesis of cell wall at multiple sites in E. coli, S. faecalis replicated new cell wall at only one site per coccus. The new wall segment was initiated and enlarged at the coccal equator, and was followed by the formation of a cross wall, centripetal growth and constriction to separate the daughter cells.


Author(s):  
Krishan Awtar

Exposure of cells to low sublethal but mitosis-arresting doses of vinblastine sulfate (Velban) results in the initial arrest of cells in mitosis followed by their subsequent return to an “interphase“-like stage. A large number of these cells reform their nuclear membranes and form large multimicronucleated cells, some containing as many as 25 or more micronuclei (1). Formation of large multinucleate cells is also caused by cytochalasin, by causing the fusion of daughter cells at the end of an otherwise .normal cell division (2). By the repetition of this process through subsequent cell divisions, large cells with 6 or more nuclei are formed.


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