Localization of the Polyadenylate Sequences in Messenger Ribonucleic Acid and in the Heterogeneous Nuclear Ribonucleic Acid of HeLa Cells

Nakazato, Hiroshi; Kopp, David W.; Edmonds, Mary

doi:10.1016/s0021-9258(19)44323-8

Cited by 107 publications

(6 citation statements)

References 24 publications

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“…The 5-28S supernatant RNA synthesized in the GH3 nuclear system resembles cellular messenger RNA in several ways. (1) It contains poly(A) tracts comparable in size to those of GH3 cytoplasmic RNA (Figure 8) and similar to those observed for a number of mRNAs from other cells (Mendecki et al, 1972;Nakazoto et al, 1973). (2) Cordycepin, a drug believed to inhibit the cytoplasmic appearance of mRNA (Adesnik et al, 1972;Penman et al, 1970), when used as the phosphorylated derivative, inhibited the extranuclear appearance of this class of supernatant RNAs (Figures 1 IB and 11C).…”

Section: Discussionsupporting

confidence: 81%

“…De novo synthesis of poly(A) by GH3 nuclei has been demonstrated (Figure 8). The poly(A) tracts isolated were comparable in size (about 150 adenine residues) to those reported for eukaryotic mRNAs (Mendecki et al, 1972;Nakazoto et al, 1973). The appearance of newly synthesized poly(A) tracts in the supernatant fraction was prevented completely by -amanitin (Figure 8B).…”

Section: Discussionsupporting

confidence: 74%

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Extended RNA synthesis in isolated nuclei from rat pituitary tumor cells

Biswas¹,

Martin²,

Tashjian³

1976

Biochemistry

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Nuclei of GH3 cells, isolated by detergent lysis, synthesized RNA for an extended period at 29 degrees C in the presence of rat liver ribonuclease inhibitor (RI). Extended RNA synthesis was dependent upon the presence of RI. Sucrose gradient sedimentation analysis of the cell-free reaction products showed that RNAs ranging from 4 S to greater than 28 S were synthesized. Further characterization of the RNA products was made by examining the sensitivity of synthesis to alpha-amanitin and actinomycin D as well as by oligo(dT)-cellulose binding properties. Evidence was obtained that RNA polymerases I, II, and III were functioning in isolated GH3 nuclei. Newly synthesized RNAs were found in both the nuclear pellet and postnuclear supernatant fractions. RNA polymerase I products remained associated with the nuclear pellet throughout a 60-min incubation period whereas RNAs synthesized by RNA polymerase III emerged rapidly into the supernatant fraction. RNA polymerase II products were distributed in both fractions and were found to contain poly(A). De novo poly(A) synthesis was demonstrated and found to be inhibited by cordvcepin triphosphate (3'-dATP). Supernatant RNAs synthesized by polymerase II contained a poly(A) segment of about 150 adenine residues; these transcripts sedimented heterogeneously with an apparent size distribution (under denaturing conditions) which was smaller than that of nuclear RNA polymerase II products and which resembled that of cellular mRNA. Qualitative differences in the nuclear and supernatant RNAs, the kinetics of appearance of the latter, and the differential effect of 3'-dATP on the extranuclear appearance of supernatant RNAs suggest that a process resembling nuclear-cytoplasmic RNA transport occurred in this cell-free nuclear system.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionsupporting

confidence: 74%

Extended RNA synthesis in isolated nuclei from rat pituitary tumor cells

Biswas¹,

Martin²,

Tashjian³

1976

Biochemistry

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“…Following this, two shorter single base sequences were detected in the HnRNA of HeLa cells. One is an internal sequence of about 25 AMPs which is transcribed rather than added posttranscriptionally as is the large poly(A) at the 3' end (Nakazato et al, 1973. The other is a poly(U) sequence of 30 to 40 nucleotides which is concentrated in the largest HnRNA molecules in regions distant from the poly(A) terminus (Molloy et al, 1972terminus (Molloy et al, , 1974.…”

mentioning

confidence: 99%

Poly(uridylic acid) sequences in messenger ribonucleic acid of HeLa cells

El¹,

Nakazato²,

Edmonds³

et al. 1976

Biochemistry

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“…Poly A preparations from ribonuclease digests of hnRNA of HeLa cells purified on oligo dT cellulose contain a small AMP-rich component which readily separates from the large poly A sequence during electrophoresis. While investigating the location of the large poly A in periodate oxidized nuclear RNA molecules which had been reduced with labeled sodium borohydride, it was noted that in contrast to large poly A, this rapidly migrating species did not become labeled (1). It was concluded that unlike large poly A, this small AMP-rich species was not at the 3' end of hnRNA molecules..…”

Section: Introductionmentioning

confidence: 99%

Properties of a small transcribed poly A sequence in heterogeneous nuclear RNA of HeLa cells

et al. 1979

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A class of heterogeneous nuclear RNA (hnRNA) molecules contain an internal transcribed poly A sequence of close to 25 uninterrupted AMP residues. HnRNA molecules containing this sequence are separable from those containing the large 3' terminal poly A sequence on the basis of their differential affinity for oligo dT cellulose. The fact that the transcribed small poly A and the 3' terminal poly A are not found in the same hnRNA molecules even though both are present in similar size classes and that the small poly A is absent from cytoplasmic messenger RNA (mRNA) has led us to propose a scheme for mRNA processing in which the 3' end of the small poly A in hnRNA becomes a priming size for the post-transcriptional addiction of the large poly A.

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Localization of the Polyadenylate Sequences in Messenger Ribonucleic Acid and in the Heterogeneous Nuclear Ribonucleic Acid of HeLa Cells

Cited by 107 publications

References 24 publications

Extended RNA synthesis in isolated nuclei from rat pituitary tumor cells

Extended RNA synthesis in isolated nuclei from rat pituitary tumor cells

Poly(uridylic acid) sequences in messenger ribonucleic acid of HeLa cells

Properties of a small transcribed poly A sequence in heterogeneous nuclear RNA of HeLa cells

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