Identification of globin mRNA in 10s RNA of rabbit reticulocytes

Kazazian, Haig H.; Moore, Patricia A.; Snyder, Pamela G.

doi:10.1016/0006-291x(73)91351-x

Cited by 15 publications

(2 citation statements)

References 14 publications

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“…Globin mRNA migrates as a single broad 9-lOS RNA band during electrophoresis in polyacrylamide gels in aqueous media (5)(6)(7)(8). However, in the presence of 98-99% formamide, polyacrylamide gel electrophoresis separates the globin mRNA into two discrete bands (8-10, 23, 24).…”

mentioning

confidence: 99%

Synthesis of DNA complementary to separated human alpha and beta globin messenger RNAs.

Forget¹,

Housman²,

Benz³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

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Human globin messenger RNA, purified by oligo(dT)-cellulose column chromatography, is reproducibly separated into two bands by polyacrylamide gel electrophoresis in the presence of 99% formamide. The more rapidly migrating (fast) band is somewhat more abundant than the slow band in normal (nonthalassemic) total reticulocyte globin messenger RNA. In a-thalassemic (Hb H disease) messenger RNA, the slow band is 6.5 times more abundant than the fast band, whereas in 0-thalassemic messenger RNA the fast band is three times more abundant than, a second band, which has a slightly greater mobility than the slow band of normal and a-thalassemic RNA. The RNA bands of nonthalassemic globin messenger RNA were eluted from the gel and efficiently transcribed into DNA copies by use of the RNA-dependent DNA polymerase of avian myeloblastosis virus. Hybridization of these copy DNAs to fast and slow band RNAs and to nonfractionated normal, a-thalassemic, and fl-thalassemic messenger RNAs revealed that the eluted fast band RNA contains predominantly a-chain specific sequences, whereas the eluted slow band RNA contains predominantly d-chain specific sequences. Nucleotide sequence analysis of 32P-labeled-RNA transcribed from the slow band copy DNA also indicated that the slow band RNA is,B messenger RNA.The ability to separate biochemically the messenger RNAs (mRNAs) for the a and ,B globin chains of human hemoglobin would greatly facilitate detailed studies of the normal human globin genes and of the precise molecular basis of the thalassemia syndromes. The a-and ,B-thalassemias are inherited disorders of human hemoglobin synthesis which are characterized by absent or decreased synthesis of a and # globin chains, respectively (1). This defect in hemoglobin synthesis has been shown, by molecular hybridization studies, to be associated with absent or decreased quantities of a or , globin chain mRNA (2-4).Globin mRNA migrates as a single broad 9-lOS RNA band during electrophoresis in polyacrylamide gels in aqueous media (5-8). However, in the presence of 98-99% formamide, polyacrylamide gel electrophoresis separates the globin mRNA into two discrete bands (8-10, 23, 24 and translated in cell-free protein synthesizing systems (8, 10, 24); the more rapid (fast) band stimulates the synthesis predominantly of a globin chains, whereas the slow band stimulates the synthesis predominantly of , globin chains.We report here the fractionation of human globin mRNA by polyacrylamide gel electrophoresis in formamide, and the characterization of its separated components by molecular hybridization and nucleotide sequencing technics utilizing DNA copies of the eluted RNA components, synthesized by means of the RNA-dependent DNA polymerase of avian myeloblastosis virus. MATERIALS AND METHODSMaterials. Most materials for RNA isolation, polyacrylamide gel electrophoresis, complementary DNA (cDNA) synthesis, and RNA-cDNA hybridization are the same as previously described (2,(11)(12)(13)

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mentioning

confidence: 99%

Synthesis of DNA complementary to separated human alpha and beta globin messenger RNAs.

Forget¹,

Housman²,

Benz³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

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“…1972). When RNA preparations are fractioned by a poly(A) selection procedure, the template activity for protein synthesis has been found to remain associated with the poly(A) containing molecules (Morrison et al 1972, Kazazian et al 1973 providing evidence for the identification of these molecules as mRNA. From the earlier studies (Fanburg et a/.…”

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confidence: 99%

Experimental Cardiac Hypertrophy and the Synthesis of Poly(A) Containing RNA and of Myocardial Proteins in the Rat: The Effect of Digitoxin Treatment

Turto

1977

Acta Physiologica Scandinavica

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The synthesis of poly(A) containing RNA was increased in heart of non-digitalized and digitalized rats after aortic constriction, the increase being of the same degree as that of the RNA lacking this sequence. No differences were found, either in the absence or presence of polyuridylic acid, in the incorporation of radioactivity into protein by cardiac ribosomes isolated from animals treated differently. It may be concluded, that after the constriction of the aorta the synthesis of mRNA proceeds at a similar rate as that of the bulk RNA, and that the treatment of the animals with digitoxin does not abolish the stimulus for hypertrophy.

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Diabetes-induced alteration in subcellular distribution of poly(A)-rich RNA from skeletal muscle

1977

Mol Cell Biochem

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Identification of globin mRNA in 10s RNA of rabbit reticulocytes

Cited by 15 publications

References 14 publications

Synthesis of DNA complementary to separated human alpha and beta globin messenger RNAs.

Synthesis of DNA complementary to separated human alpha and beta globin messenger RNAs.

Experimental Cardiac Hypertrophy and the Synthesis of Poly(A) Containing RNA and of Myocardial Proteins in the Rat: The Effect of Digitoxin Treatment

Diabetes-induced alteration in subcellular distribution of poly(A)-rich RNA from skeletal muscle

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