Autoradiographic study of deoxyribonucleic acid synthesis in L cells infected with the agent of meningopneumonitis

Moore, Dorothy E. (University of Chicago, Chicago, Ill.), and James W. Moulder. Autoradiographic study of deoxyribonucleic acid synthesis in L cells infected with the agent of meningopneumonitis. J. Bacteriol. 92:1128-1132. 1966.-L cells infected with the agent of meningopneumonitis were labeled with H(3)-cytidine at 5-hr intervals after infection, and cell samples were fixed every 5 hr after labeling. These preparations were then digested with ribonuclease, stained by the Feulgen procedure, and examined by autoradiography. Labeled meningopneumonitis inclusions were first seen 15 hr after infection. Deoxyribonucleic acid (DNA) was synthesized in both L-cell nuclei and meningopneumonitis agent for as long as 40 hr after infection. Nuclear DNA synthesis was unaffected until 25 hr after infection, at which time synthesis of agent DNA reached its peak. After 25 hr, both meningopneumonitis and L cell DNA synthesis declined.     

Phagostimulation of larval Culexpipiens L. by nucleic acid nucleotides, nucleosides and bases

ABSTRACT. The influence of nucleic acid constituents on the rate of ingestion of charcoal powder filtered from ambient water by larvae of Culex pipiens L. was examined. All nucleotides tested stimulated ingestion to some extent. Various mono-, di- and tri-phosphates of adenosine were most effective and at concentrations of 1 mM stimulated ingestion nearly as well as yeast extract, a powerful phagostimu-lant. Guanylic, thymidylic, cytidylic and uridylic acids were less stimulatory, the latter two even at 10 mM. Cyclic AMP and deoxyadenylic acid were less effective than other adenine nucleotides. The nucleosides adenosine, guanosine and uridine were almost as effective as their corresponding nucleotides (adenylic, guanylic and uridylic acids); thymidine was less effective than thymidylic acids, whereas cytidine was non-stimulatory. Adenine, guanine, uracil and cytosine, the bases of the ribose nucleotides, were non-phagostimulatory, whereas thymine, base of the deoxynucleotide, thymidylic acid, caused low but significantly increased ingestion. These findings are compared with the reported phagostimulation by nucleic acid constituents of certain plant feeding insects and with the stimulation of engorgement of the blood meal by many blood feeding insects.     

Transport of nucleic acid bases and nucleosides across the bacterial membrane

Abstract Defects have been found in a colorimetric assay developed by its inventors for determining rapidly and quantitatively whether strains of Escherichia coli are resistant to the lethality of mammalian serum. Results gained with the colorimetric assay on the serum resistance of E. coli strains bearing colicin V plasmids differed from results gained with an assay in which the rate of growth of the bacterial strains in a medium containing rabbit serum was estimated turbidimetrically. The colorimetric assay was intended by its developers to facilitate studies of the genetics of serum resistance in pathogenic bacteria. When used as originally described, the assay is likely to mislead in such studies.     

Uptake in vitro of nucleic acid precursors and nucleic acids by Zajdela ascitic hepatoma cells.

Zajdela ascitic hepatoma cells are shown to take up pyrimidine bases at much lower rates than obtained in slices from normal rat liver. The rates of uptake of adenine and uridine by the Zajdela cells are, however, as high as in the slices. Like the slices, again, the Zajdela cells take up E. coli RNA and DNA at very low rates but, unlike the slices, thses cells degrade rapidly the RNA taken up. The Zajdela cells resemble parenchymal cell suspensions derived from normal rat liver in regard to the uptake of pyrimidine bases and the ability to degrade heterologous RNA.     

Mutagenicity of irradiated solutions of nucleic acid bases and nucleosides in Salmonella typhimurium

Solutions of nucleic acid bases, nucleosides and a nucleotide, saturated with either N2, N2O or O2, were irradiated and tested for mutagenicity towards Salmonella typhimurium, with and without pre-incubation. Irradiated solutions of the nucleic acid bases were all non-mutagenic. Irradiated solutions of the nucleosides showed mutagenicity in S. typhimurium TA100 (pre-incubation assay). Generally, the mutagenicity followed the order: N2O greater than N2 greater than O2. The results show that the formation of mutagenic radiolytic products is initiated by attack of mainly OH radicals on the 2-deoxy-D-ribose moiety of the nucleosides. With irradiated solutions of the nucleotide, thymidine-5'-monophosphate, no mutagenicity could be detected.     

Affinity chromatography of nucleosides and nucleic acid base derivatives with nucleic acid bases or nitrobenzeneboronic acid substituted silicas

Nucleic acid bases such as adenine and uracil, and nitrobenzeneboronic acid substituted silicas were prepared by the reaction of chloromethylbenzene substituted silica with adenine sodium salt and trimethylsilylated uracil, and nitration of benzeneboronic acid substituted silica, respectively. From the results of HPLC of nucleosides and N-ethyl derivatives of nucleic acid bases using modified silicas, hydrophobic base stacking interaction, selective hydrogen bonding interaction between purine and pyrimidine bases, and reversible cyclic boronate ester formation between diols of nucleosides with boronic acid were effective for the separation of nucleic acid related compounds. Moreover, association constants for hydrogen bonding formation of nucleic acid bases were estimated.     

Use of automated chromatography on the amino acid analyzer with lithium citrate buffers to separate nucleic acid bases, nucleosides, nucleotides and their precursors

A simple and sensitive method has been developed to separate nucleic acid bases, nucleosides, nucleotides and their precursors by automated chromatography using the amino acid analyzer with lithium citrate buffers. The method is sensitive to a concentration of 5 nmol, linear in the range of 5--100 nmol, and resolves almost all the bases, nucleosides, nucleotides and their precursors of physiologic importance.     

Nucleic acid-like structures II. Polynucleotide analogues as possible primitive precursors of nucleic acids

Activated derivatives of purine-containing deoxynucleoside- diphosphates spontaneously oligomerize to produce pyrophosphate- linked oligodeoxynucleotide analogues. These analogues are of potential interest as models of primitive, polynucleotide precursors. The efficiency of oligomerization (ImpdGpIm and ImpdApIm much greater than ImpdIpIm) appears to reflect a combination of stacking forces and the specific geometric orientations of the stacked units. Under favorable conditions, chain lengths greater than 20 have been obtained for oligomers containing pdGp in the absence of a template. In the presence of a complementary template, the activated derivatives of pdGp and pdAp oligomerize much more extensively. An acyclo-analogue of G has also been shown to undergo template-directed oligomerization on poly(C). These observations suggest the possibility that primitive information transfer might have evolved in much simpler systems and that this function was taken over by polynucleotides at a later stage in evolution. For the previous paper in this series see Schwartz and Orgel, 1985a.     

Differences in incorporation of nucleic acid bases and nucleosides by various Mycoplasma and Acholeplasma species.

Eight species representative of the serological diversity of the Mycoplasmatales were tested for their ability to incorporate radiolabeled nucleic acid precursors into acid-insoluble material. Cultures in complex growth medium were centrifuged and resuspended in minimal essential medium (Eagle). For Acholeplasma laidlawii, labeling occurred mainly during the first 4 h of incubation, with substrate saturation at 20 micron. All organisms tested incorporated uracil, adenine, and guanine; none incorporated cytosine. Thymine was incorporated only by bovine group 7, Mycoplasma putrefaciens, and Mycoplasma pneumoniae (strain 3546), but deoxynucleosides enhanced thymine incorporation in A. laidlawii, Mycoplasma gallisepticum, M. pneumoniae (strain AP-164), and Mycoplasma hyorhinis. Nucleoside incorporation (adenosine, guanosine, uridine, cytidine, and thymidine) was not observed for the arginine-utilizing species, Mycoplasma hominis and Mycoplasma arginini, whereas all other organisms tested incorporated nucleosides. The incorporation pattern provides additional metabolic evidence to support the biochemical and antigenic diversity of these organisms. The recognition of differences in incorporation of nucleic acid precursors is important not only to the specific labeling of these organisms, but also to the study of metabolism and transport.