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Discovery of Nuclear DNA-like RNA (dRNA, hnRNA) and Ribonucleoproteins Particles Containing hnRNA.

Georgiev GP - Acta Naturae (2016 Jan-Mar)

Bottom Line: Therefore, I was invited to represent Russia at the Symposium and deliver a speech on these topics.However, my visa had only been issued after the end of the Symposium, and, therefore, the presentation was delivered by my former colleague G.N.The transcript of the lecture is presented below.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, 119334, Russia.

ABSTRACT
On August 9-11, 2014, Cold Spring Harbor (USA) hosted a special symposium dedicated to the discovery of messenger or informational RNA and the main events in the subsequent studies of its synthesis, regulation of synthesis, maturation, and transport. The existence of mRNA in bacteria was first suggested in 1961 by Jacob and Monod, based on genetic studies [1]. The same year, Brenner et al. confirmed the hypothesis [2]. Our laboratory played a key role in the discovery of messenger RNA in eukaryotes, as well as in the discovery of the nuclear ribonucleoproteins that contain it and in the elucidation of their structural organization. Therefore, I was invited to represent Russia at the Symposium and deliver a speech on these topics. However, my visa had only been issued after the end of the Symposium, and, therefore, the presentation was delivered by my former colleague G.N. Yenikolopov, who works at Cold Spring Harbor Laboratory. The transcript of the lecture is presented below.

No MeSH data available.


Related in: MedlinePlus

Isolation and properties of “phenolic” cell nuclei. (left panel) Scheme ofcell nuclei isolation by phenol treatment. A photograph of the “phenolic” nucleiof Ehrlich ascites carcinoma cells is presented. (right-hand panel) Compositionof the obtained nuclei and properties of their RNA: intermediate nucleotidecomposition between those of DNA and rRNA; ultracentrifugation data.
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Figure 1: Isolation and properties of “phenolic” cell nuclei. (left panel) Scheme ofcell nuclei isolation by phenol treatment. A photograph of the “phenolic” nucleiof Ehrlich ascites carcinoma cells is presented. (right-hand panel) Compositionof the obtained nuclei and properties of their RNA: intermediate nucleotidecomposition between those of DNA and rRNA; ultracentrifugation data.

Mentions: My main collaborator in the discovery of nuclear dRNA was V.L. Mantieva, whowent on to earn a PhD in biology. We were interested in the nature of nuclearRNA [3] and used a newly developed phenolmethod to isolate RNA from cells [4]. Asuspension of mouse Ehrlich ascites carcinoma cells was shaken in 0.14 M NaCland phenol at pH 6.0 and 4°C, followed by centrifugation. Surprisingly, inaddition to the expected aqueous and phenol phases, the centrifugation producedan intermediate layer that contained cell nuclei that retained their shape[5]. These nuclei contained chromatin andnucleoli, which stored DNA, nuclear RNA, and most of the nuclear proteins(Fig. 1).Since phenol inhibits enzyme activity, we believedthat “phenolic” nuclei can be a good source of nuclear RNA. Later,it was shown that nuclear RNA can indeed be extracted from“phenolic” nuclei by this procedure if it is performed at 65°C[3]. The isolated nuclear RNA containedcomponents with sedimentation coefficients of 28S and 18S, typical forribosomal RNA, and heterogeneous material. The nucleotide composition of thenuclear RNA was intermediate between mouse DNA (G+C/A+T = 0.72) and ribosomalRNA (G+C/A+U = 1.65)(Fig. 1).It seemed that nuclear RNAcontained ribosomal RNA and a new type of RNA whose nucleotide composition wassimilar to that of DNA: i.e., informational RNA. The first experiments on thefractionation of nuclear RNA, conducted in 1961, confirmed this hypothesis[3].


Discovery of Nuclear DNA-like RNA (dRNA, hnRNA) and Ribonucleoproteins Particles Containing hnRNA.

Georgiev GP - Acta Naturae (2016 Jan-Mar)

Isolation and properties of “phenolic” cell nuclei. (left panel) Scheme ofcell nuclei isolation by phenol treatment. A photograph of the “phenolic” nucleiof Ehrlich ascites carcinoma cells is presented. (right-hand panel) Compositionof the obtained nuclei and properties of their RNA: intermediate nucleotidecomposition between those of DNA and rRNA; ultracentrifugation data.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837567&req=5

Figure 1: Isolation and properties of “phenolic” cell nuclei. (left panel) Scheme ofcell nuclei isolation by phenol treatment. A photograph of the “phenolic” nucleiof Ehrlich ascites carcinoma cells is presented. (right-hand panel) Compositionof the obtained nuclei and properties of their RNA: intermediate nucleotidecomposition between those of DNA and rRNA; ultracentrifugation data.
Mentions: My main collaborator in the discovery of nuclear dRNA was V.L. Mantieva, whowent on to earn a PhD in biology. We were interested in the nature of nuclearRNA [3] and used a newly developed phenolmethod to isolate RNA from cells [4]. Asuspension of mouse Ehrlich ascites carcinoma cells was shaken in 0.14 M NaCland phenol at pH 6.0 and 4°C, followed by centrifugation. Surprisingly, inaddition to the expected aqueous and phenol phases, the centrifugation producedan intermediate layer that contained cell nuclei that retained their shape[5]. These nuclei contained chromatin andnucleoli, which stored DNA, nuclear RNA, and most of the nuclear proteins(Fig. 1).Since phenol inhibits enzyme activity, we believedthat “phenolic” nuclei can be a good source of nuclear RNA. Later,it was shown that nuclear RNA can indeed be extracted from“phenolic” nuclei by this procedure if it is performed at 65°C[3]. The isolated nuclear RNA containedcomponents with sedimentation coefficients of 28S and 18S, typical forribosomal RNA, and heterogeneous material. The nucleotide composition of thenuclear RNA was intermediate between mouse DNA (G+C/A+T = 0.72) and ribosomalRNA (G+C/A+U = 1.65)(Fig. 1).It seemed that nuclear RNAcontained ribosomal RNA and a new type of RNA whose nucleotide composition wassimilar to that of DNA: i.e., informational RNA. The first experiments on thefractionation of nuclear RNA, conducted in 1961, confirmed this hypothesis[3].

Bottom Line: Therefore, I was invited to represent Russia at the Symposium and deliver a speech on these topics.However, my visa had only been issued after the end of the Symposium, and, therefore, the presentation was delivered by my former colleague G.N.The transcript of the lecture is presented below.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gene Biology, Russian Academy of Sciences, Vavilova Str., 34/5, Moscow, 119334, Russia.

ABSTRACT
On August 9-11, 2014, Cold Spring Harbor (USA) hosted a special symposium dedicated to the discovery of messenger or informational RNA and the main events in the subsequent studies of its synthesis, regulation of synthesis, maturation, and transport. The existence of mRNA in bacteria was first suggested in 1961 by Jacob and Monod, based on genetic studies [1]. The same year, Brenner et al. confirmed the hypothesis [2]. Our laboratory played a key role in the discovery of messenger RNA in eukaryotes, as well as in the discovery of the nuclear ribonucleoproteins that contain it and in the elucidation of their structural organization. Therefore, I was invited to represent Russia at the Symposium and deliver a speech on these topics. However, my visa had only been issued after the end of the Symposium, and, therefore, the presentation was delivered by my former colleague G.N. Yenikolopov, who works at Cold Spring Harbor Laboratory. The transcript of the lecture is presented below.

No MeSH data available.


Related in: MedlinePlus