Abstract
We have adapted the oocyte injection procedure for the detection of regulatory components involved in the transcription of a eukaryotic mRNA gene. Injection of the histone gene repeat h22 DNA of Psammechinus miliaris into the Xenopus oocyte nucleus results in correct initiation of the histone mRNAs, but readthrough by RNA polymerase occurs at the 3' end of the H3 histone gene (Hentschel, C. C., Probst, E. & Birnstiel, M. L. (1980) Nature (London) 288, 100-102). Coinjection into the oocyte of a chromosomal salt wash fraction derived from sea urchin embryos results in the generation of authentic 3' termini of the histone H3 mRNA. We have partially purified the protein component by column chromatography and density gradient centrifugation. The regulatory factor binds to heparin columns and, hence, has the properties anticipated of an RNA- or DNA-binding protein. The sedimentation coefficient of the active component was determined to be about 12 S, suggesting a molecular weight of 200,000-250,000.
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- Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
- Birchmeier C., Grosschedl R., Birnstiel M. L. Generation of authentic 3' termini of an H2A mRNA in vivo is dependent on a short inverted DNA repeat and on spacer sequences. Cell. 1982 Apr;28(4):739–745. doi: 10.1016/0092-8674(82)90053-8. [DOI] [PubMed] [Google Scholar]
- Chatterjee S., Goldstein L. Proteins in nucleocytoplasmic interactions. V. Intranuclear localization of proteins in Amoeba proteus. J Cell Biol. 1971 Jan;48(1):202–207. doi: 10.1083/jcb.48.1.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Robertis E. M., Lienhard S., Parisot R. F. Intracellular transport of microinjected 5S and small nuclear RNAs. Nature. 1982 Feb 18;295(5850):572–577. doi: 10.1038/295572a0. [DOI] [PubMed] [Google Scholar]
- Diaz M. O., Barsacchi-Pilone G., Mahon K. A., Gall J. G. Transcripts from both strands of a satellite DNA occur on lampbrush chromosome loops of the newt Notophthalmus. Cell. 1981 Jun;24(3):649–659. doi: 10.1016/0092-8674(81)90091-x. [DOI] [PubMed] [Google Scholar]
- Grosschedl R., Birnstiel M. L. Delimitation of far upstream sequences required for maximal in vitro transcription of an H2A histone gene. Proc Natl Acad Sci U S A. 1982 Jan;79(2):297–301. doi: 10.1073/pnas.79.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grosschedl R., Birnstiel M. L. Spacer DNA sequences upstream of the T-A-T-A-A-A-T-A sequence are essential for promotion of H2A histone gene transcription in vivo. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7102–7106. doi: 10.1073/pnas.77.12.7102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hentschel C. C., Birnstiel M. L. The organization and expression of histone gene families. Cell. 1981 Aug;25(2):301–313. doi: 10.1016/0092-8674(81)90048-9. [DOI] [PubMed] [Google Scholar]
- Hentschel C., Probst E., Birnstiel M. L. Transcriptional fidelity of histone genes injected into Xenopus oocyte nuclei. Nature. 1980 Nov 6;288(5786):100–102. doi: 10.1038/288100a0. [DOI] [PubMed] [Google Scholar]
- Kedes L. H., Birnstiel M. L. Reiteration and clustering of DNA sequences complementary to histone messenger RNA. Nat New Biol. 1971 Apr 7;230(14):165–169. doi: 10.1038/newbio230165a0. [DOI] [PubMed] [Google Scholar]
- Kressmann A., Clarkson S. G., Telford J. L., Birnstiel M. L. Transcription of xenopus tDNAmet1 and sea urchin histone DNA injected into the Xenopus oocyte nucleus. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):1077–1082. doi: 10.1101/sqb.1978.042.01.108. [DOI] [PubMed] [Google Scholar]
- Levy S., Childs G., Kedes L. Sea urchin nuclei use RNA polymerase II to transcribe discrete histone RNAs larger than messengers. Cell. 1978 Sep;15(1):151–162. doi: 10.1016/0092-8674(78)90091-0. [DOI] [PubMed] [Google Scholar]
- Platt T. Termination of transcription and its regulation in the tryptophan operon of E. coli. Cell. 1981 Apr;24(1):10–23. doi: 10.1016/0092-8674(81)90496-7. [DOI] [PubMed] [Google Scholar]
- Probst E., Kressmann A., Birnstiel M. L. Expression of sea urchin histone genes in the oocyte of Xenopus laevis. J Mol Biol. 1979 Dec 15;135(3):709–732. doi: 10.1016/0022-2836(79)90173-6. [DOI] [PubMed] [Google Scholar]
- Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
- Schaffner W., Gross K., Telford J., Birnstiel M. Molecular analysis of the histone gene cluster of psammechinus miliaris: II. The arrangement of the five histone-coding and spacer sequences. Cell. 1976 Aug;8(4):471–478. doi: 10.1016/0092-8674(76)90214-2. [DOI] [PubMed] [Google Scholar]
- Spelsberg T. C., Hnilica L. S. Deoxyribonucleoproteins and the tissue-specific restriction of the deoxyribonucleic acid in chromatin. Biochem J. 1970 Nov;120(2):435–437. doi: 10.1042/bj1200435. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stunnenberg H. G., Wennekes L. M., van den Broek W. H. RNA polymerase from the fungus, Aspergillus nidulans. Large-scale purification of DNA-dependent RNA polymerase I (or A). Eur J Biochem. 1979 Jul;98(1):107–119. doi: 10.1111/j.1432-1033.1979.tb13167.x. [DOI] [PubMed] [Google Scholar]
- Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weinberg E. S., Birnstiel M. L., Purdom I. F., Williamson R. Genes coding for polysomal 9S RNA of sea urchins: conservation and divergence. Nature. 1972 Nov 24;240(5378):225–228. doi: 10.1038/240225a0. [DOI] [PubMed] [Google Scholar]