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D W Li

Publications and source records attributed to D W Li.

22 records · Page 2Linked to original sources

Mouse thymidylate synthase messenger RNA lacks a 3' untranslated region.

Analysis of the sequence of cDNA corresponding to mouse thymidylate synthase (5,10-methylenetetrahydrofolate:dUMP C-methyltransferase, EC 2.1.1.45) mRNA revealed that the termination codon TAA was followed immediately by a poly(A) sequence. This raised the possibility that mouse thymidylate synthase mRNA lacks a 3' untranslated region. In the present study, we have further investigated this possibility. DNA corresponding to the 3' end of the thymidylate synthase gene was isolated from a genomic library. The sequence of the genomic DNA was identical to that of the cDNA in the coding region. However, the termination codon was TAG in the genomic sequence rather than TAA, and poly(A) was not present in the genomic DNA. Sequences flanking the site of poly(A) addition were in good agreement with polyadenylylation consensus sequences. S1 nuclease analysis revealed that approximately 80% of the thymidylate synthase mRNA molecules were polyadenylylated at the termination codon. A secondary polyadenylylation site was detected 190-200 nucleotides downstream of the primary site. We conclude that the major species of mouse thymidylate synthase mRNA lacks a 3' untranslated region and that the final A of the termination codon is added by poly(A) polymerase. It appears that a 3' untranslated region is not essential for the accumulation or translation of this mRNA.

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Extensive homology of nuclear ribonucleic acid and polysomal poly(adenylic acid) messenger ribonucleic acid between normal and neoplastically transformed cells.

A cell line, designated BP6T, derived from Syrian hamster embryo (SHE) cells following treatment with benzo[a]pyrene is capable of producing tumors in newborn hamsters following the injection of as few as 1-10 cells. Polysomal poly(A) mRNA and total nuclear RNA obtained from this highly tumorigenic cell line were compared to RNAs obtained from the nonneoplastic parental embryo cells by a variety of techniques. RNA excess hybridizations to normal cell radiolabeled single-copy DNA or to a single-copy DNA tracer enriched for sequences transcribed in neoplastically transformed cells were unable to detect any significant differences in RNA sequence complexity between normal SHE cells and neoplastic BP6T cells. This finding of extensive homology of polysomal poly(A) mRNA and total nuclear RNA between normal and neoplastic cells, together with our previous finding of extensive homology of the major 35S-labeled nuclear or cytoplasmic polypeptides observable on two-dimensional gels [Leavitt, J. C., & Moyzis, R. K. (1978) J. Biol. Chem. 253, 2497-2500], demonstrates that the phenotypic changes associated with neoplastic transformation by chemical carcinogens are accompanied by relatively few changes in the qualitative pattern of gene expression in cells cultured in vitro.

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