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. 1984 Feb;3(2):473–479. doi: 10.1002/j.1460-2075.1984.tb01831.x

Localization of small nuclear polymerase I RNA sequences at the 5' end of the human rDNA transcription unit.

R Reichel, B J Benecke
PMCID: PMC557369  PMID: 6325172

Abstract

A human ribosomal DNA clone isolated from a genomic library was used to localize the DNA sequences coding for HeLa cell small nuclear polymerase I RNA (snPI RNA). By using a subcloned 1.2-kb EcoRI-SalI fragment, including the initiation region of the 45S rRNA transcription unit, it was shown that the snPI RNA sequences are located within the first 600 nucleotides of the 5' end of the external transcribed spacer. Strand-specific hybridization following exonuclease III digestion of the plasmid containing the 1.2-kb Eco-Sal fragment demonstrated that the snPI RNA molecules and the 45S pre-rRNA are transcribed from the same coding strand. A detailed mapping of individual snPI RNA molecules showed that most of these small RNA species span the putative early processing site at position 415 of the external transcribed spacer of the human rRNA precursor.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benecke B. J., Penman S. A new class of small nuclear RNA molecules synthesized by a type I RNA polymerase in HeLa cells. Cell. 1977 Dec;12(4):939–946. doi: 10.1016/0092-8674(77)90158-1. [DOI] [PubMed] [Google Scholar]
  2. Benecke B. J., Penman S. Large species differences in the pattern of snPI RNA which can distinguish ape from human. J Cell Biol. 1979 Mar;80(3):778–783. doi: 10.1083/jcb.80.3.778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  4. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  5. Financsek I., Mizumoto K., Mishima Y., Muramatsu M. Human ribosomal RNA gene: nucleotide sequence of the transcription initiation region and comparison of three mammalian genes. Proc Natl Acad Sci U S A. 1982 May;79(10):3092–3096. doi: 10.1073/pnas.79.10.3092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kafatos F. C., Jones C. W., Efstratiadis A. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979 Nov 24;7(6):1541–1552. doi: 10.1093/nar/7.6.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kohorn B. D., Rae P. M. Localization of DNA sequences promoting RNA polymerase I activity in Drosophila. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3265–3268. doi: 10.1073/pnas.80.11.3265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
  9. Miesfeld R., Arnheim N. Identification of the in vivo and in vitro origin of transcription in human rDNA. Nucleic Acids Res. 1982 Jul 10;10(13):3933–3949. doi: 10.1093/nar/10.13.3933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Miller J. R., Hayward D. C., Glover D. M. Transcription of the 'non-transcribed' spacer of Drosophila melanogaster rDNA. Nucleic Acids Res. 1983 Jan 11;11(1):11–19. doi: 10.1093/nar/11.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moss T. A transcriptional function for the repetitive ribosomal spacer in Xenopus laevis. Nature. 1983 Mar 17;302(5905):223–228. doi: 10.1038/302223a0. [DOI] [PubMed] [Google Scholar]
  12. Murphy J. T., Burgess R. R., Dahlberg J. E., Lund E. Transcription of a gene for human U1 small nuclear RNA. Cell. 1982 May;29(1):265–274. doi: 10.1016/0092-8674(82)90111-8. [DOI] [PubMed] [Google Scholar]
  13. Reichel R., Benecke B. J. Reinitiation of synthesis of small cytoplasmic RNA species K and L in isolated HeLa cell nuclei in vitro. Nucleic Acids Res. 1980 Jan 25;8(2):225–234. doi: 10.1093/nar/8.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reichel R., Monstein H. J., Jansen H. W., Philipson L., Benecke B. J. Small nuclear RNAs are encoded in the nontranscribed region of ribosomal spacer DNA. Proc Natl Acad Sci U S A. 1982 May;79(10):3106–3110. doi: 10.1073/pnas.79.10.3106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roop D. R., Kristo P., Stumph W. E., Tsai M. J., O'Malley B. W. Structure and expression of a chicken gene coding for U1 RNA. Cell. 1981 Mar;23(3):671–680. doi: 10.1016/0092-8674(81)90430-x. [DOI] [PubMed] [Google Scholar]
  16. Rungger D., Achermann H., Crippa M. Transcription of spacer sequences in genes coding for ribosomal RNA in Xenopus cells. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3957–3961. doi: 10.1073/pnas.76.8.3957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scheer U., Trendelenburg M. F., Krohne G., Franke W. W. Lengths and patterns of transcriptional units in the amplified nucleoli of oocytes of Xenopus laevis. Chromosoma. 1977 Mar 16;60(2):147–167. doi: 10.1007/BF00288462. [DOI] [PubMed] [Google Scholar]
  18. Smith A. J. The use of exonuclease III for preparing single stranded DNA for use as a template in the chain terminator sequencing method. Nucleic Acids Res. 1979 Mar;6(3):831–848. doi: 10.1093/nar/6.3.831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  21. Wellauer P. K., Dawid I. B. Isolation and sequence organization of human ribosomal DNA. J Mol Biol. 1979 Mar 5;128(3):289–303. doi: 10.1016/0022-2836(79)90089-5. [DOI] [PubMed] [Google Scholar]
  22. Yanofsky C. Prokaryotic mechanisms in eukaryotes? Nature. 1983 Apr 28;302(5911):751–752. doi: 10.1038/302751a0. [DOI] [PubMed] [Google Scholar]
  23. Zieve G., Benecke B. J., Penman S. Synthesis of two classes of small RNA species in vivo and in vitro. Biochemistry. 1977 Oct 4;16(20):4520–4525. doi: 10.1021/bi00639a029. [DOI] [PubMed] [Google Scholar]
  24. Zylber E. A., Penman S. Products of RNA polymerases in HeLa cell nuclei. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2861–2865. doi: 10.1073/pnas.68.11.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]

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