Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1973 Jul;70(7):2029–2032. doi: 10.1073/pnas.70.7.2029

Synthesis of Globin Ribonucleic Acid from Duck-Reticulocyte Chromatin In Vitro

Richard Axel 1, Howard Cedar 1, Gary Felsenfeld 1
PMCID: PMC433658  PMID: 4516203

Abstract

The proteins of chromatin serve to restrict the transcription of DNA. The relevance of these findings to the control of gene expression is contingent upon the demonstration that this restriction is specific and mirrors the patterns of RNA synthesis observed in vivo. In this study we demonstrate by RNA-DNA hybridization that the vast majority of the chromatin-directed RNA is synthesized from the unique regions of the reticulocyte genome. Furthermore, by use of the DNA complement of globin mRNA as a probe in annealing reactions, de novo synthesis of globin RNA was detected in RNA transcripts from duck reticulocyte chromatin. No globin sequences were detected in similar preparations of RNA in vitro either from liver chromatin or from DNA freed of protein. These results show that the proteins of chromatin serve to restrict transcription in a very specific manner and provide convincing evidence for the existence of transcriptional control factors in eukaryotes.

Keywords: DNA transcription, RNA-DNA hybridization

Full text

PDF
2030

Selected References

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

  1. Attardi G., Parnas H., Hwang M. I., Attardi B. Giant-size rapidly labeled nuclear ribonucleic acid and cytoplasmic messenger ribonucleic acid in immature duck erythrocytes. J Mol Biol. 1966 Sep;20(1):145–182. doi: 10.1016/0022-2836(66)90123-9. [DOI] [PubMed] [Google Scholar]
  2. Axel R., Schlom J., Spiegelman S. Evidence for translation of viral-specific RNA in cells of a mouse mammary carcinoma. Proc Natl Acad Sci U S A. 1972 Mar;69(3):535–538. doi: 10.1073/pnas.69.3.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bank A., Terada M., Metafora S., Dow L., Marks P. A. In vitro synthesis of DNA components of human genes for globins. Nat New Biol. 1972 Feb 9;235(58):167–169. doi: 10.1038/newbio235167a0. [DOI] [PubMed] [Google Scholar]
  4. Bekhor I., Kung G. M., Bonner J. Sequence-specific interaction of DNA and chromosomal protein. J Mol Biol. 1969 Jan;39(2):351–364. doi: 10.1016/0022-2836(69)90322-2. [DOI] [PubMed] [Google Scholar]
  5. Bonner J., Dahmus M. E., Fambrough D., Huang R. C., Marushige K., Tuan D. Y. The Biology of Isolated Chromatin: Chromosomes, biologically active in the test tube, provide a powerful tool for the study of gene action. Science. 1968 Jan 5;159(3810):47–56. doi: 10.1126/science.159.3810.47. [DOI] [PubMed] [Google Scholar]
  6. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  7. Kacian D. L., Watson K. F., Burny A., Spiegelman S. Purification of the DNA polymerase of avian myeloblastosis virus. Biochim Biophys Acta. 1971 Sep 24;246(3):365–383. doi: 10.1016/0005-2787(71)90773-8. [DOI] [PubMed] [Google Scholar]
  8. McConaughy B. L., McCarthy B. J. Fractionation of chromatin by thermal chromatography. Biochemistry. 1972 Mar 14;11(6):998–1003. doi: 10.1021/bi00756a008. [DOI] [PubMed] [Google Scholar]
  9. Melli M., Whitfield C., Rao K. V., Richardson M., Bishop J. O. DNA-RNA hybridization in vast DNA excess. Nat New Biol. 1971 May 5;231(18):8–12. [PubMed] [Google Scholar]
  10. Paul J., Gilmour R. S. Organ-specific restriction of transcription in mammalian chromatin. J Mol Biol. 1968 Jul 14;34(2):305–316. doi: 10.1016/0022-2836(68)90255-6. [DOI] [PubMed] [Google Scholar]
  11. Ross J., Aviv H., Scolnick E., Leder P. In vitro synthesis of DNA complementary to purified rabbit globin mRNA (RNA-dependent DNA polymerase-reticulocyte-hemoglobin-density gradient centrifugation-oligo(dT) primer). Proc Natl Acad Sci U S A. 1972 Jan;69(1):264–268. doi: 10.1073/pnas.69.1.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Schutz G., Beato M., Feigelson P. Isolation of eukaryotic messenger RNA on cellulose and its translation in vitro. Biochem Biophys Res Commun. 1972 Nov 1;49(3):680–689. doi: 10.1016/0006-291x(72)90465-2. [DOI] [PubMed] [Google Scholar]
  13. Simpson R. T., Sober H. A. Circular dichroism of calf liver nucleohistone. Biochemistry. 1970 Aug 4;9(16):3103–3109. doi: 10.1021/bi00818a001. [DOI] [PubMed] [Google Scholar]
  14. Verma I. M., Temple G. F., Fan H., Baltimore D. In vitro synthesis of DNA complementary to rabbit reticulocyte 10S RNA. Nat New Biol. 1972 Feb 9;235(58):163–167. doi: 10.1038/newbio235163a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES