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G Dennert

Publications and source records attributed to G Dennert.

116 records · Page 7Linked to original sources

Effects of oleate starvation in a fatty acid auxotroph of Escherichia coli K-12.

The effects of oleate starvation on an oleate auxotroph of Escherichia coli K-12 were investigated. Following removal of oleate from the mutant growing in a minimal glycerol-peptone medium, the cells stopped making deoxyribonucleic acid, ribonucleic acid, protein, and phospholipids; they began to die exponentially and finally lysed. During oleate starvation in minimal medium minus peptone, inhibition of macromolecular syntheses and death occurred; however, lysis did not follow. When growth ceased, no further dying was observed. It is shown that none of the early effects (inhibition of macromolecular syntheses and death) can be due to leakiness of the cells, induction of a prophage or a colicin, or lack of energy sources. The cause of inhibition of macromolecular syntheses remained unknown. Since the rate of death was the same as the generation time under different conditions, it appears that death is due to the defective synthesis of some cellular structure (quite possibly, cytoplasmic membrane) during phospholipid deficiency. Lysis was found to require protein synthesis; electron microscopy revealed a peculiar type of "lysis from within"; i.e., the shape of the cells did not change but fragmentation of the inner layer of the cell envelope occurred. The murein was found to be unaltered. Most likely, lysis was a consequence of the cell's attempt to synthesize cytoplasmic membrane with altered phospholipid composition or during phospholipid deficiency. Several membrane functions (respiration, adenosine triphosphate formation, permeability) existing before oleate removal were not lost during starvation. Therefore, general damage to the membrane did not occur, and it could be that most, if not all, described effects were due to defective de novo membrane synthesis.

Adenosine Triphosphate↗

CTX-B inhibits CTL cytotoxicity and cytoskeletal movements.

Binding of cytotoxic T lymphocytes (CTL) to specific targets induces cytoskeletal movements in the effector cell followed by delivery of the lethal hit which ultimately results in target cell lysis. The question whether movement of the cytoskeleton in CTL are obligatory for delivery of the lethal hit is not resolved. Here we report that the CTX-B subunit of cholera toxin which is devoid of the catalytic CTX-A subunit inhibits CTL function. Inhibition was found not to be due to interference with TCR expression, CTL-target conjugate formation, target induced transmembrane signalling or secretion of BLT-esterase. CTX-B however does interfere with F-actin patch formation at the effector target binding site and inhibits reorientation of the microtubule organizing center and Golgi apparatus towards the target binding site. It is concluded that interference with cytoskeletal movements is responsible for inhibition of cytolysis pointing to an important role of the cytoskeleton in the lytic reaction.

Actins↗

Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells.

The formation of ultrastructural membrane lesions of varying size during cell mediated cytolysis effected by human peripheral blood leukocytes was recently reported by Dourmashkin et al. and Henkart et al. Using cloned mouse natural killer (NK) cells as effectors and YAC-1 cells or rabbit erythrocytes as targets, we now report two types of membrane lesions with inner diameters of 16 +/- 2 nm and approximately 5 nm, respectively. These lesions arise by membrane insertion of tubular complexes that may be assembled from subunits during the cytolytic reaction. The tubules are detected on target membranes by immune electron microscopy and appear to form transmembrane channels as seen in ultrathin sections. Both tubules are partially purified by membrane extraction with SDS and gel filtration in deoxycholate containing buffer. Based on the correlation of tubule assembly and cytolysis and on their detection on target membranes, we suggest that both types of tubules may be related to cytolysis.

Cell Membrane↗