B McStay — Page 2 | PubMed Health

PubMed1983

The origin of the rRNA precursor from Xenopus borealis, analysed in vivo and in vitro.

We have determined the origin of the major transcript of Xenopus borealis rDNA by the use of an SI nuclease protection assay. The DNA surrounding the origin of this transcript was sequenced, and the region upstream of the origin was shown to have strong sequence homology with that region from X.laevis rDNA. We have also demonstrated faithful transcription from this origin using cloned X.borealis rDNA in an extract derived from X. laevis culture cells. This in vitro transcription was insensitive to 100 micrograms/ml alpha-amanatin, suggesting that it was mediated by RNA polymerase 1.

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PubMed1983

DNaseI-hypersensitive sites at promoter-like sequences in the spacer of Xenopus laevis and Xenopus borealis ribosomal DNA.

We have detected a DNAseI hypersensitive site in the ribosomal DNA spacer of Xenopus laevis and Xenopus borealis. The site is present in blood and embryonic nuclei of each species. In interspecies hybrids, however, the site is absent in unexpressed borealis rDNA, but is present normally in expressed laevis rDNA. Hypersensitive sites are located well upstream (over lkb) of the pre-ribosomal RNA promoter. Sequencing of the hypersensitive region in borealis rDNA, however, shows extensive homology with the promoter sequence, and with the hypersensitive region in X. laevis. Of two promoter-like duplications in each spacer, only the most upstream copy is associated with hypersensitivity to DNAaseI. Unlike DNAaseI, Endo R. MspI digests the rDNA of laevis blood nuclei at a domain extending downstream from the hypersensitive site to near the 40S promoter. Since the organisation of conserved sequence elements within this "proximal domain" is similar in three Xenopus species whose spacers have otherwise evolved rapidly, we conclude that this domain plays an important role in rDNA function.

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