Production of superoxide anion and hydrogen peroxide by KB cells in an anoxia-reoxygenation model, and role of allopurinol.
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To investigate the metabolic fate of a 6S RNA (VA-RNA), which appears in human cell lines only after infection with adenovirus, various species of RNA from the cells at different stages of infection were tested for the presence of nucleotide sequence homologous to this RNA. RNA extracted from subvellular fractions of the cells was further fractionated by sucrose gradient centrifugation and analysed by acrylamide gel electrophoresis or by competition against 32P-labeled VA-RNA in hybridization with adenovirus DNA. The results pointed to the following conclusions. (1) VA-RNA is synthesized in the cells throughout the entire period after infections, starting before the onset of viral DNA synthesis. (2) VA-RNA does not participate in protein synthesis as a messenger RNA or a precursor of transfer RNA. (3) The nucleotide sequence homologous to VA-RNA exists as a part of certain species of nuclear high molecular weight RNA in the infected cells.
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The immunoelectrophhoretic analysis of two preparations of the same virus cultivated in different substrates (Sendai/egg and Sendai/KB) demonstrate that virus envelopes contain: a) identical virus-specific antigens (but also some virus antigens with only partial immunochemical identity, or even non-identity); b) antigens incorporated by the virus from the host cell, during assembly and release. The latter differ according to the host, and some of them are only partially identical with the corresponding antigens of the host cell, which suggests that host cell glycoproteins are incorporated into the virus envelope after certain virus-induced modifications.
Metaphase chromosomes and interphase nuclei have been isolated by two different procedures, using either acidic or alkaline media. The morphological integrity of the isolated material has been monitored by electron microscopy. In the acidic procedure the thickness of the chromosome fibers is reduced but the chromosome gross structure is preserved. However, electrophoretic analysis indicates complete disappearance of histones H1 and partial loss of H2A and H2B. No significant modification of acid-soluble proteins isolated from chromosomes or nuclei was observed after 5-BrdU incorporation for 72 hours.
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