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S M Anderson

Publications and source records attributed to S M Anderson.

At least 145 records · Page 8Linked to original sources

Identification of the viral sequence required for the generation of recovered avian sarcoma viruses and characterization of a series of replication-defective recovered avian sarcoma viruses.

The ability of transformation-defective deletion mutants of Schmidt-Ruppin Rous sarcoma virus to induce tumors and generate recovered sarcoma viruses (rASVs) was correlated with the partial src sequences retained in the transformation-defective viral genomes. Since all the transformation-defective viruses that were capable of generating rASVs retained a portion of the 3' src sequence, regardless of the extent of the 5' src deletion, and those lacking the 3' src were unable to generate rASVs, it appears that the 3', but most likely not the 5', src sequence retained in the transformation-defective viral genome is essential for rASV formation. However, rASVs derived from a particular mutant, td109, which retained a portion of the 3' src sequence, but lacked most (if not all) of the 5' src sequence, were all found to be defective in replication. Analyses of the genomic sequences of 13 isolates of td109-derived rASVs revealed that they contained various deletions in viral envelope (env), polymerase (pol), and structural protein (gag) genes. Ten isolates of rASVs contained env deletions. One isolate (rASV3812) contained a deletion of env and the 3' half of pol, and one isolate (rASV398) contained a deletion of env and pol. The one with the most extensive deletion (rASV374) had a deletion from the p12-coding sequence through pol and env. In addition, the 5' src region of td109-derived rASVs were heterogeneous. Among the 7 isolates analyzed in detail, one isolate of rASV had a small deletion of the 5' src sequence, whereas three other isolates contained extra new sequences upstream from src. Both env- and env- pol- rASVs were capable of directing the synthesis of precursor and mature gag proteins in the infected nonproducer cells. We attribute the deletions in the replication-defective rASVs to the possibility that the 5' recombination site between the td109 and c-src sequence, involved regions of only partial homology due to lack of sufficient 5' src sequence in the td109 genome for homologous recombination. A model of recombination between the viral genome and the c-src sequence is proposed to account for the requirement of the 3' src sequence and the basis for the generation of deletions in td109-derived rASVs.

Animals↗

The measurement of monoamine oxidase activity: a comparison of the ion exchange resin column method and the solvent extraction method.

A comparison was made of monoamine oxidase (MAO) activity level; the level was obtained by quantification of radiolabeled deaminated metabolites separated from unchanged amines by two different methods. MAO activity in whole brain of male and female mice from five inbred strains was measured with three substrates PEA, TYR, and 5-HT to examine apparent variations in enzyme activity level with substrate metabolized. Levels of activity measured by the two separation methods were equivalent, as demonstrated by comparison of population mean activity levels and by Pearson product-moment correlation coefficients (0.92, 0.93, and 0.83 for PEA, TYR, and 5-HT, respectively).

Animals↗

Biochemical and molecular analysis of naturally occurring Adh variants in Drosophila melanogaster.

The results of a detailed analysis of the biochemical and molecular basis of alcohol dehydrogenase (ADH) activity variation existing among six naturally occurring and one ethyl methanesulfonate-induced Adh variant strain of Drosophila melanogaster are presented. Significant specific activity differences exist among the strains but the majority of ADH activity variation can be accounted for by differences in levels of ADH protein. These protein level differences can, in turn, be accounted for by ADH synthesis rate variation, which positively correlates with in vivo levels of cytoplasmic Adh mRNA. The functional variability is correlated with known structural variation in and around the area of the Adh gene.

Alcohol Dehydrogenase↗

Construction and isolation of a transforming murine retrovirus containing the src gene of Rous sarcoma virus.

Recombinant murine retroviruses containing the src gene of the avian retrovirus Rous sarcoma virus were isolated. Such viruses were isolated from cells after transfection with DNAs in which the src gene was inserted into the genome of the amphotropic murine retrovirus 4070A. The isolated viruses had functional gag and pol genes, but they were all env defective since the src gene was inserted in the middle of the env gene coding region. Infectious transforming virus could be isolated only from cells transfected with DNA constructions in which the src gene was in the same polarity as that of a long terminal repeat of the amphotropic viral genome. These recombinant viruses encoded a pp60src protein with a molecular weight similar to that of the Schmidt-Ruppin strain of Rous sarcoma virus. In addition, the src protein(s) of these recombinant viruses was as active as protein kinases in the immune complex protein kinase assay. Intravenous injection of helper-independent Moloney and Friend murine leukemia virus pseudotypes of the src recombinant viruses into 6-week-old NIH Swiss mice resulted in the appearance of splenic foci within 2 weeks, splenomegaly and, later after infection (8 to 10 weeks), anemia. Infectious transforming virus could be recovered from the spleens of diseased animals. Such viruses encoded pp60src but not p21ras or mink cell focus-forming virus-related glycoproteins.

Animals↗

Interactions of erythrosin B (U.S. F, D & C Red 3) with rat cortical membranes.

Erythrosin B inhibits Na,K-ATPase in rat brain tissue as demonstrated by studying glycoside binding, ATPase activity and ion fluxes. The potency of the noncompetitive inhibition of [3H]-ouabain binding by erythrosin B is influenced by glycoside concentration, monovalent cation concentration, and incubation time. [14C]-Erythrosin B binds to synaptic membranes prepared from rat cortex. Erythrosin B and some of its structural analogs inhibit both [3H]-ouabain and [14C]-erythrosin B binding, but ouabain and other glycosides do not inhibit the binding of [14C]-erythrosin B. Subcellular distributions of [3H]-ouabain and [14C]-erythrosin B binding in fractionated cortical tissue preparations are equivalent and parallel ATPase activity. The dissimilar response of [3H]-ouabain binding and [14C]-erythrosin B binding to changes in tissue preparation, incubation temperature, and partial solubilization of binding sites by deoxycholate (DOC) suggests two separate binding sites for erythrosin and ouabain to rat cortical membranes.

Animals↗

Effect of environmental alcohol on in vivo properties of Drosophila alcohol dehydrogenase.

The effects of environmental 2-propanol on the in vivo properties of Drosophila alcohol dehydrogenase (E.C. 1..1.1.1.) are presented. Exposed flies were found to exhibit a significant decrease in ADH specific activity with a concomitant increase in the enzyme's relative in vivo stability and concentration. The possible adaptive significance of the observed responses is discussed.

1-Propanol↗

Ethanol consumption: selective breeding in mice.

From a foundation stock of HS/Ibg mice, mass selection has yielded high-ethanol acceptance (HEA) and low-ethanol acceptance (LEA) lines with distributions which show little overlap in the 10th generation. Divergence has been systematic and the estimate of the realized heritability after 10 generations of selective breeding is 0.21 +/- 0.04. Low to moderate correlations among three methods of measuring alcohol consumption, including the one on which selection was based, were shown in an F2 generation derived from C57BL/Ibg and C3H/Ibg mice.

Alcohol Drinking↗

Identification of strychnine binding sites in the rat retina.

[3H]Strychnine specifically binds to membrane fractions isolated from rat retinae. The binding is saturable, with an apparent dissociation constant, KD, of 14.3 x 10(-9) M and 205 fmol bound/mg protein. Specific binding is time-dependent and proportional to protein concentration. Glycine and taurine are equally potent inhibitors of [3H]strychnine binding (Ki = 4 x 10(-5) M); no other amino acids endogenously present in the retina inhibited [3H]strychnine binding.

Amino Acids↗

In vitro translation of avian myeloblastosis virus RNA.

Avian myeloblastosis virus (AMV)-infected cells contain two viral mRNA's, a genome-sized 34S (7.5-kilobase) mRNA and a 21S (2.5-kilobase) subgenomic mRNA, which contains the AMV-specific sequences (myb sequences). We found that AMV virions packaged both the 7.5-kilobase full-length genomic RNA and the 2.5-kilobase subgenomic RNA. In vitro translation of AMV virion RNA sized by sucrose density gradient centrifugation yielded 76,000-, 56,000-, 48,500-, 47,000-, and 32,000-dalton products. The 76,000-dalton protein was coded for by RNA throughout the gradient, but the peak of activity was at 34S to 35S. The 56,000-, and 48,500-, and 32,000-dalton proteins were encoded in a 21S RNA, and 47,000-dalton protein was encoded in an RNA of approximately 24S. The 76,000-dalton protein was identified as Pr76gag, based upon immunoprecipitation with specific antiserum and the presence of the 19* dipeptide. 7-Methylguanosine triphosphate inhibited the syntheses of Pr76gag and the 56,000-, 48,500-, and 32,000-dalton proteins, but not the synthesis of the 47,000-dalton protein. The 56,000-, 48,500-, 47,000-, and 32,000-dalton proteins were not immunoprecipitated by anti-gag, anti-reverse transcriptase, or anti-gp85 antiserum. Two-dimensional peptide maps of the 56,000- and 48,500-dalton proteins indicated that they were unique. In vitro translational products of myeloblastosis-associated virus 1 were also analyzed to aid in the identification of the AMV myb gene product(s); the translational products analyzed included Pr76gag, p60env, and a 56,000-dalton polypeptide which apparently was not identical to the 56,000-dalton AMV translational product, as determined by two-dimensional peptide mapping. Our data indicated that one of these proteins (56,000, 48,500, or 32,000 daltons) may represent the product of the AMV myb gene and, therefore, the putative transforming protein(s) of AMV.

Amino Acid Sequence↗

Changes in levels of alcohol dehydrogenase during the development of Drosophila melanogaster.

The results of an analysis of the biochemical basis of changes in alcohol dehydrogenase (E.C.1.1.1.1) activity over Drosophila development are presented. The data indicate that (1) the characteristic changes that occur in ADH activity over development are predominantly, it not exclusively, the result of quantitative changes in the amount of enzyme present rather than qualitative changes affecting the enzyme's specific activity and (2) the fluctuations in amount of ADH which occur during development are not the result of the only known form of post-translational modification capable of affecting the biochemical properties of the enzyme. We conclude that developmental changes in amount of ADH are most likely the result of fluctuations in the turnover of the ADH protein.

Alcohol Oxidoreductases↗

Characterization of the transforming gene of Fujinami sarcoma virus.

The src gene present in all avian sarcoma viruses is not present in the genome of Fujinami sarcoma virus, a potent sarcoma-inducing virus in chickens. Fujinami virus is defective and requires helper virus for replication. RNA from a mixture of helper and transforming viruses consists of two components, 35S and 28S. Oligonucleotide fingerprinting of each RNA component revealed that the 35S component was identical to the RNA of the helper virus. Thus, the genome of Fujinami virus must be the 28S RNA, which corresponds approximately to a molecular weight of 1.7 x 10(6) or 5300 nucleotides. Fujinami viral RNA shares several oligonucleotides with helper viral RNA at both 3' and 5' ends but contains a unique sequence of at least 3000 nucleotides in the middle of the genome. Fujinami viral RNA contains no src-specific oligonucleotides of the Rous sarcoma virus genome and did not hybridize with DNA complementary to the src sequences. The 60,000-dalton src protein of Rous sarcoma virus was undetectable in Fujinami virus-transformed cells. Instead, these transformed cells contain a protein of 140,000 daltons precipitable by antisera against virion proteins, which is likely to be the transforming protein of this virus.

Alpharetrovirus↗

Biochemical differences between products of the Adh locus in Drosophila.

An analysis of the molecular properties of the major alcohol dehydrogenase (E.C.1.1.1.1.) allozyme variants found segregating in natural populations of D. melanogaster is presented. Our results indicate: (1) ADH-S enzyme has generally lower Michaelis-Menten constants than those of ADH-F; (2) ADH-S and ADH-F enzymes display opposite interactions for both co-factor and substrate; and (3) higher levels of ADH are associated with the Adh-fast genotype. The possible adaptive significance of these findings is discussed.

Alcohol Oxidoreductases↗

Avian erythroblastosis virus produces two mRNA's.

We analyzed the viral mRNA's present in fibroblast nonproducer clones transformed by avian erythroblastosis virus. Two size classes of mRNA (28 to 30S and 22 to 24S) were identified by solution hybridization with both complementary DNA strong stop and complementary DNA made against the unique sequences of avian erythroblastosis virus. Based upon the kinetics of hybridization with complementary DNA made against the unique sequences of avian erythroblastosis virus, we estimated that there were 400 to 500 copies of the 28 to 30S RNA per cell and 200 to 250 copies of the 22 to 24S RNA per cell. Both RNA species were packaged in the virion. In vitro translation of the 28 to 30S virion RNA yielded a 75,000-dalton protein which was the 75,000-dalton gag-related polyprotein found in avian erythroblastosis virus-transformed cells. In vitro translation of the 22 to 24S virion RNA yielded two proteins (46,000 and 48,000 daltons). This indicates that there may be two genes in avian erythroblastosis virus, one coding for the 75,000-dalton gag-related polyprotein and the second coding for the 46,000- or 48,000-dalton protein or both.

Alpharetrovirus↗

Ethanol consumption and hepatic enzyme activity.

Enzyme activity and ethanol consumption were measured in an F2 generation derived from the C57BL and C3H inbred mouse strains. A significant correlation (0.25) was found between alcohol dehydrogenase activity and ethanol acceptance in the F2 generation. Mass selection from a genetically heterogenous mouse stock, HS/Ibg, has yielded high ethanol acceptance (HEA) and low ethanol acceptance (LEA) lines of mice. The mean ethanol acceptance scores for the fifth generation of these lines are 1.008 and 0.606, respectively. The total liver alcohol dehydrogenase activity was found to be 24% higher in the HEA line than in the LEA line after five generations of selective breeding. No association between cytosolic aldehyde dehydrogenase activity and ethanol acceptance was found in either the F2 generation or the fifth generation of the selectively bred lines.

Alcohol Drinking↗