Dataset Information

Differential sensitivity of RSVts (temperature-sensitive Rous-sarcoma virus)-infected rat kidney cells to nucleoside antibiotics at permissive and non-permissive temperatures.

ABSTRACT: Among a variety of anti-tumour agents tested, oxanosine and 5-azacytidine were found to be significantly more effective in inhibiting growth of rat kidney cells infected with a temperature-sensitive mutant of Rous sarcoma virus at a permissive temperature (33 degrees C) than at a non-permissive temperature (39 degrees C). These two nucleoside antibiotics were antagonistic to each other in cytotoxicity. They seem to share the same carrier-mediated membrane-transport system, because dipyridamole, a potent inhibitor of nucleoside transport, protected cells from the cytotoxicity of both drugs. Thymidine transport, which is twice as fast in cells at 33 degrees C as at 39 degrees C, was competitively inhibited by both drugs. Thus the differential toxicity of oxanosine and 5-azacytidine at the two temperatures is thought to be due to their increased transport via the thymidine-transport system, which is somehow under the influence of the active src-gene product.

SUBMITTER: Uehara Y

PROVIDER: S-EPMC1152957 | biostudies-other | 1985 Dec

REPOSITORIES: biostudies-other

ACCESS DATA

Similar Datasets

Project description:UnlabelledPurified retroviral Gag proteins can assemble in vitro to form immature virus-like particles (VLPs). By cryoelectron tomography, Rous sarcoma virus VLPs show an organized hexameric lattice consisting chiefly of the capsid (CA) domain, with periodic stalk-like densities below the lattice. We hypothesize that the structure represented by these densities is formed by amino acid residues immediately downstream of the folded CA, namely, the short spacer peptide SP, along with a dozen flanking residues. These 24 residues comprise the SP assembly (SPA) domain, and we propose that neighboring SPA units in a Gag hexamer coalesce to form a six-helix bundle. Using in vitro assembly, alanine scanning mutagenesis, and biophysical analyses, we have further characterized the structure and function of SPA. Most of the amino acid residues in SPA could not be mutated individually without abrogating assembly, with the exception of a few residues near the N and C termini, as well as three hydrophilic residues within SPA. We interpret these results to mean that the amino acids that do not tolerate mutations contribute to higher-order structures in VLPs. Hydrogen-deuterium exchange analyses of unassembled Gag compared that of assembled VLPs showed strong protection at the SPA region, consistent with a higher-order structure. Circular dichroism revealed that a 29mer SPA peptide shifts from a random coil to a helix in a concentration-dependent manner. Analytical ultracentrifugation showed concentration-dependent self-association of the peptide into a hexamer. Taken together, these results provide strong evidence for the formation of a critical six-helix bundle in Gag assembly.ImportanceThe structure of a retrovirus like HIV is created by several thousand molecules of the viral Gag protein, which assemble to form the known hexagonal protein lattice in the virus particle. How the Gag proteins pack together in the lattice is incompletely understood. A short segment of Gag known to be critical for proper assembly has been hypothesized to form a six-helix bundle, which may be the nucleating event that leads to lattice formation. The experiments reported here, using the avian Rous sarcoma virus as a model system, further define the nature of this segment of Gag, show that it is in a higher-order structure in the virus particle, and provide the first direct evidence that it forms a six-helix bundle in retrovirus assembly. Such knowledge may provide underpinnings for the development of antiretroviral drugs that interfere with virus assembly.

Dataset Information

Differential sensitivity of RSVts (temperature-sensitive Rous-sarcoma virus)-infected rat kidney cells to nucleoside antibiotics at permissive and non-permissive temperatures.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets