ABSTRACT
The expression of Xenopus laevis larval and adult globin genes after phenylhydrazine-induced anemia has been investigated at the cellular and molecular levels by means of cloned cDNA probes specific for the four main larval and the four main adult globin mRNA species. In the circulating blood of anemic metamorphic larvae there are at least two distinct populations of erythroblasts containing either larval or adult globin mRNA sequences. The cells expressing adult sequences replace those expressing larval ones at the end of metamorphosis. Under the influence of anemia the qualitative pattern of transcribed RNA species is not changed, but the larval to adult transition takes place earlier during development. This would imply that this transition is not strongly correlated with the morphological changes of metamorphosis. The abundance of the different larval globin mRNA species is similar and, as compared to control animals, not affected by the phenylhydrazine treatment. In anemic adults no reactivation of larval gene expression has been observed. The different adult globin mRNA species are present in comparable abundance, but the phenylhydrazine treatment appears to enhance expression of the alpha A II and beta A II genes, as compared to nonanemic individuals.
Subject(s)
Erythroblasts/physiology , Globins/genetics , RNA, Messenger/genetics , Xenopus laevis/genetics , Anemia/genetics , Animals , Cloning, Molecular , Gene Expression Regulation , Metamorphosis, Biological , Nucleic Acid Hybridization , Xenopus laevis/growth & developmentABSTRACT
Purified vitellogenin mRNA of Xenopus laevis was incubated with mechanically sheared DNA in high concentrations of formamide and the resulting R-loops (i.e. RNA . DNA hybrid fragments) separated from the bulk DNA by caesium chloride buoyant density centrifugation. Hybridization with 125I-labeled vitellogenin mRNA revealed a 15--30-fold enrichment of the DNA coding for vitellogenin. Restriction analysis of the R-loop-enriched DNA demonstrated that all known endonuclease HindIII fragments coding for vitellogenin of unfractionated Xenopus DNA were also present in the enriched material, including the specific fragments for the oligo(A)-containing segment of the RNA. Comparison of these restriction data with the structure found in cloned vitellogenin cDNA, indicates the presence of at least one intervening sequence in the genomic DNA coding for vitellogenin.
