Cytoplasmic synthesis of globin RNA in differentiated murine erythroleukemia cells: possible involvement of RNA-dependent RNA polymerase
Three lines of evidence indicate that RNA-dependent RNA synthesis occurs in mouse erythroleukemia cells. The first involves labeling studies with (/sup 3/H)uridine and shows a greater initial labeling rate of globin RNA in the cytoplasm than in the nucleus. Labeled globin RNA found in the cytoplasm after a very short pulse with tritiated uridine is of the mature 9S size while labeled globin RNA in the nuclei is exclusively in the form of 15S precursor molecules, suggesting that cytoplasmic globin RNA is not of nuclear origin. A high concentration of actinomycin D has no effect on the initial rate of labeling of cytoplasmic globin RNA, supporting this conclusion. Other experiments showed that the labeling of cytoplasmic globin RNA does not involve end addition to preexisting globin RNA. The second line of evidence is the identification of globin RNA minus strand in the cytoplasm of differentiated murine erythroleukemia cells by hybridization with single-stranded DNA probes containing the strand of the same sense as globin mRNA. This material has the same electrophoretic mobility as globin RNA and hybridizes with probes containing only the 5' part or only the 3' part of the gene suggesting that it is a full size copy of globin RNA. Finally, in murine erythroleukemia cells an RNA-dependent RNA polymerase activity is detected by using poly(A) oligo(U) as a template-primer combination. This activity increases significantly after induction, suggesting that it is differentiation specific.
- Research Organization:
- Boston Biomedical Research Institute, MA
- OSTI ID:
- 5402394
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:5
- Country of Publication:
- United States
- Language:
- English
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MESSENGER-RNA
BIOSYNTHESIS
LABELLING
RNA POLYMERASES
BIOCHEMISTRY
GLOBIN
HYBRIDIZATION
MICE
TRITIUM COMPOUNDS
TUMOR CELLS
URIDINE
ANIMAL CELLS
ANIMALS
AZINES
CHEMISTRY
ENZYMES
HETEROCYCLIC COMPOUNDS
HYDROXY COMPOUNDS
LABELLED COMPOUNDS
MAMMALS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
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PYRIMIDINES
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RNA
RODENTS
SYNTHESIS
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URACILS
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550201* - Biochemistry- Tracer Techniques