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Biomedical subjects

V G Allfrey

Publications and source records attributed to V G Allfrey.

At least 91 records · Page 5Linked to original sources

Selective synthesis and accumulation of nuclear non-histone proteins during carcinogenesis of the colon induced by 1, 2-dimethylhydrazine.

The complement of nuclear non-histone proteins in epithelial cells of the colon is progressively altered during the course of carcinogenesis induced by 1, 2-dimethylhydrazine, until finally the nuclear proteins of tumor cells are easily distinguishable from those of the surrounding normal tissue. These changes in nuclear protein compostion reflect earlier differences in the rates of synthesis of individual protein species. Radioisotopic double-labeling experiments show that the synthesis of nuclear proteins of molecular weights 44,000 and 62,000 is selectively accelerated within 4 weeks after administration of the carcinogen, long before any morphological indications of malignancy appear.

Amino Acids↗

Selective effects of inhibitors of protein synthesis on metabolism of nuclear anc cytoplasmic proteins: evidence for coordinate synthesis of non-histone chromosomal proteins.

We have compared the effects of inhibitors of protein synthesis on the metabolism of nuclear and cytoplasmic proteins of HeLa S-3 cells. L-1-tosylamido-2-phenylethyl chloromethyl ketone, a potent inhibitor of polypeptide chain initiation, was shown to preferentially inhibit the synthesis of cytoplasmic proteins and of histones at concentrations that permit continued amino acid incorporation into nuclear non-histone proteins. Comparisons of the molecular weight distributions of newly synthesized proteins in the presence and absence of L-1-tosylamido-2-phenylethyl chloromethyl ketone have revealed striking differences between nuclear anc cytoplasmic protein fractions. Differential effects on the synthesis of cytoplasmic proteins, acid-soluble nuclear proteins, and residual nuclear proteins have also been obtained with the antibiotic, pactamycin, another inhibitor of polypeptide chain initiation. The incorporation of radioactive amino acids into nuclear non-histone proteins shows resistance to inhibition by pactamycin, but is strongly inhibited by agents such as puromycin and cycloheximide which block chain elongation. The possibility that proliferating cells have developed specialized mechanisms for the coordinate synthesis of chromosomal proteins, possibly involving polycistronic messenger RNAs, is tested and discussed.

Cell Nucleus↗

Fractionation of nuclei and analysis of nuclear proteins of rat liver and Morris hepatoma 7777.

The contributions of nuclear populations to the total profile of nuclear proteins in a tissue were examined in normal rat liver and Morris hepatoma 7777. Comparison by sodium dodecyl sulfate polyacrylamide gel electrophoresis of phenol-soluble nuclear proteins from tumor and control liver revealed additional proteins of molecular weight 60,000, 100,00, and 135,000 and the loss of proteins of about 45,000 and 55,000 in the tumor. Subfractionation of liver nuclei on a 30 to 50% sucrose gradient yielded three nuclear classes with nearly identical complements of the phenol-soluble proteins. Similar fractionation performed on the hepatoma nuclei also produced three nuclear populations. In the hepatoma nuclei, several differences in the phenol-soluble proteins were found between the minor, slowly sedimenting nuclear fraction, and the two major fractions, while the two latter fractions were very similar in their protein composition. Histones derived from both tissues were also compared electrophoretically, indicating a decrease in the concentration of histone H1(0)in all nuclear classes derived from the tumor.

Animals↗

Cell-free translation of the globin message within polydisperse high-molecular-weight ribonucleic acid of avian erythrocytes.

Polydisperse high-molecular-weight RNA of nucleated avian erythrocytes includes sequences coding for globin chains. The RNA was extracted from immature erythrocytes of ducks and fractionated under denaturing conditions by sucrose density gradient centrifugation in 99% dimethylsulfoxide. The RNA sedimenting faster than 45 S was able to direct the synthesis of duck globins in the Krebs II ascites cell-free protein-synthesizing system. The newly synthesized globin molecules have been identified by their characteristic electrophoretic properties in polyacrylamide gels containing either urea or sodium dodecyl sulfate, and by immunoprecipitation of the released globin chains by rabbit antibodies against duck hemoglobin. In order to rule out the possibility of a contamination of the high-molecular-weight RNA with duck-globin messenger RNA tailing from the 9-10S region, rabbit-globin messenger RNA was added to duck RNA as an internal control. No rabbit-globin messenger RNA activity could be detected in the RNA fractions sedimenting faster than 45 S. It is concluded that high-molecular-weight RNAs in the nucleated erythroid cell contain sequences of globin messenger RNAs covalently attached to larger polynucleotide chains. These results support the view that polydisperse nuclear RNA is the precursor of the cytoplasmic messenger RNA fraction.

Animals↗