1976
DOI: 10.1111/j.1432-1033.1976.tb10580.x
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The Use of Mercurated Nucleoside Triphosphate as a Probe in Transcription Studies in vitro

Abstract: 1. Purified form A RNA polymerase and the endogenous, nuclear form A RNA polymerase are shown to incorporate 5-mercuri-uridine 5'-triphosphate (Hg-UTP) into RNA in vitro. The K , for both Hg-UTP and UTP are in the region of 10 pM for the purified enzyme.2. The RNA products formed in nucleoli by endogenous RNA polymerase A have similar base compositions (G + C-rich) whether UTP or Hg-UTP is provided as the substrate in vitro.3. Sulphydryl-Sepharose chromatography of RNA synthesised in vitro by nucleoli allows s… Show more

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Cited by 27 publications
(20 citation statements)
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“…In vitro synthesis in isolated nuclei or chromatin was carried out in the presence of 5-mercuriuridine triphosphate (Hg-UTP). The rate of RNA synthesis is essentially the same either in the presence of Hg-UTP or ordinary U T P as measured by incorporation of [3H]GTP in accord with previous observations by other investigators (Dale et al, 1973;Smith and Huang, 1976;Crouse et al, 1976;Beebee and Butterworth, 1976;Biessmann et al, 1976). Addition of saturating amounts of E .…”
Section: In Vivo Content Of Viral Sequences In Pulse-labeled Rnasupporting
confidence: 82%
See 1 more Smart Citation
“…In vitro synthesis in isolated nuclei or chromatin was carried out in the presence of 5-mercuriuridine triphosphate (Hg-UTP). The rate of RNA synthesis is essentially the same either in the presence of Hg-UTP or ordinary U T P as measured by incorporation of [3H]GTP in accord with previous observations by other investigators (Dale et al, 1973;Smith and Huang, 1976;Crouse et al, 1976;Beebee and Butterworth, 1976;Biessmann et al, 1976). Addition of saturating amounts of E .…”
Section: In Vivo Content Of Viral Sequences In Pulse-labeled Rnasupporting
confidence: 82%
“…Many of these studies suggest that chromatin and nuclei appear to retain their in vivo template specificities in transcription of specific genes (Axel et al, 1973;Gilmour and Paul, 1973;Astrin, 1973;Shih et al, 1973;Steggles et al, 1974;Rymo et al, 1974;Jacquet et al, 1974;Wilson et al, 1975;Gilmour et al, 1975;Marzluff and Huang, 1975;Stein et al, 1975;Tsai et al, 1976). Recent reports on the use of mercurated nucleotide triphosphate as the substrate for RNA synthesis in isolated nuclei or chromatin and purification of the Hg-tagged RNA by sulfhydryl affinity column have been met with interest (Smith and Huang, 1976;Crouse et al, 1976;Beebee and Butterworth, 1976;Biessmann et al, 1976). It seems to offer a novel method of resolving the newly synthesized in vitro RNA from any preexisting nuclear and chromatin RNA, and to permit the use of specific radioactive cDNA probes to analyze the in vitro products.…”
mentioning
confidence: 97%
“…RESULTS Binding of Mercurated Nucleic Acids to SH-Agarose. In order to follow specific transcription in isolated nuclei, we distinguished in vitro synthesized from preexisting mRNA sequences either by incorporation of a mercurated nucleotide (9,(16)(17)(18)(19) and subsequent isolation of the mercurated RNA by affinity chromatography on SH-agarose or by incorporation of a radioactive nucleotide and isolation of the specific hybrids after hybridization to mercurated cDNA. The basis of both procedures is the affinity of mercurated polynucleotides for SH-agarose.…”
Section: Methodsmentioning
confidence: 99%
“…According to Beebee and Butterworth and Zasloff and Felsenfeld inadequate hybridisation is obtained with RNA containing the level of mercuration chosen here, i.e. 25 % substitution in a single nucleotide [38,49]. However, the decrease in rate of hybridisation observed by those authors is explicable entirely in terms of the absence of a suitable thiol ligand for the hgRNA.…”
Section: Discussionmentioning
confidence: 93%