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. 1973 May;70(5):1355–1358. doi: 10.1073/pnas.70.5.1355

Sequence Homology at the 5′-Termini of Insect Messenger RNAs

Joseph Ilan 1, Judith Ilan 1
PMCID: PMC433496  PMID: 4351173

Abstract

Treatment of insect polyribosomes with 1 M KCl released a messenger ribonucleoprotein with a pronounced 16S peak. Phenol extraction resulted in a defined peak of 10S RNA, which was judged as mRNA by the following criteria: it showed specificity for binding to ribosomes, and the formation of initiation complex was dependent on protein initiation factors, GTP, mRNA, and aminoacyl-tRNA. The complex directed protein synthesis upon the addition of elongation factors. mRNA was treated with phosphatase and phosphorylated at the 5′-end with [32P]cyanoethylphosphate. [32P]mRNA was digested by T1 ribonuclease to completion and chromatographed on DEAE-cellulose. The only fragment with 32P was 15 nucleotides long; it was treated with pancreatic ribonuclease and fingerprinted. Fractions of AC, AAC, and AAAC were found. Initiation signal AUG or GUG in these mRNAs does not begin immediately at the 5′-end and may be at a distance greater than 15 nucleotides. Alkaline hydrolysis of mRNAs labeled in vivo with [14C]adenosine revealed Ap and pppAp. Alkaline hydrolysis of mRNA labeled with 32P at the 5′-terminus resulted in pAp. Hence, these results suggest that in a heterogeneous population of mRNAs from insects, all start with A and have sequence homology at the 5′-termini. This sequence may reflect the signal for RNA polymerase on the gene or may promote the binding of mRNA to ribosomes.

Keywords: Tenebrio, poly(A), initiation, chromatography, fingerprinting

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Selected References

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

  1. Adams J. M., Cory S. Untranslated nucleotide sequence at the 5'-end of R17 bacteriophage RNA. Nature. 1970 Aug 8;227(5258):570–574. doi: 10.1038/227570a0. [DOI] [PubMed] [Google Scholar]
  2. Brown J. C., Smith A. E. Initiator codons in eukaryotes. Nature. 1970 May 16;226(5246):610–612. doi: 10.1038/226610a0. [DOI] [PubMed] [Google Scholar]
  3. Burr H., Lingrel J. B. Poly A sequences at the 3' termini of rabbit globin mRNAs. Nat New Biol. 1971 Sep 8;233(36):41–43. doi: 10.1038/newbio233041a0. [DOI] [PubMed] [Google Scholar]
  4. Darnell J. E., Wall R., Tushinski R. J. An adenylic acid-rich sequence in messenger RNA of HeLa cells and its possible relationship to reiterated sites in DNA. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1321–1325. doi: 10.1073/pnas.68.6.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. De Wachter R., Fiers W. Sequences at the 5'-terminus of bacteriophage Q-beta-RNA. Nature. 1969 Jan 18;221(5177):233–235. doi: 10.1038/221233a0. [DOI] [PubMed] [Google Scholar]
  6. DeWachter R., Verhassel J. P., Fiers W. The 5'-terminal and group of the RNA of the bacteriophage MS2. Biochim Biophys Acta. 1968 Mar 18;157(1):195–197. [PubMed] [Google Scholar]
  7. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Heywood S. M. Specificity of mRNA binding factor in eukaryotes. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1782–1788. doi: 10.1073/pnas.67.4.1782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hsu W. T., Weiss S. B. Selective translation of T4 template RNA by ribosomes from T4-infected Escherichia coli. Proc Natl Acad Sci U S A. 1969 Sep;64(1):345–351. doi: 10.1073/pnas.64.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ilan J., Ilan J. A possible role of the AUG codon in the initiation of polypeptide synthesis in a eukaryotic orgamism. Biochim Biophys Acta. 1970 Dec 14;224(2):614–619. doi: 10.1016/0005-2787(70)90594-0. [DOI] [PubMed] [Google Scholar]
  11. Ilan J., Ilan J. An mRNA bound initiation factor and its role in translation of natural message. Nat New Biol. 1973 Feb 7;241(110):176–180. doi: 10.1038/newbio241176a0. [DOI] [PubMed] [Google Scholar]
  12. Ilan J., Ilan J. Stage-specific initiation factors for protein synthesis during insect development. Dev Biol. 1971 Jun;25(2):280–292. doi: 10.1016/0012-1606(71)90031-5. [DOI] [PubMed] [Google Scholar]
  13. Ilan J. Studies on template-active ribonucleic acid from nuclei of insect pupae. Biochemistry. 1969 Dec;8(12):4825–4831. doi: 10.1021/bi00840a026. [DOI] [PubMed] [Google Scholar]
  14. Ilan J., Tokuyasu K. Synthesis of arabinosyl-ATP by photophosphorylation. Life Sci II. 1971 Jun 8;10(11):637–642. doi: 10.1016/0024-3205(71)90059-2. [DOI] [PubMed] [Google Scholar]
  15. Labrie F. Isolation of an RNA with the properties of haemoglobin messenger. Nature. 1969 Mar 29;221(5187):1217–1222. doi: 10.1038/2211217a0. [DOI] [PubMed] [Google Scholar]
  16. Lee S. Y., Mendecki J., Brawerman G. A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cells. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1331–1335. doi: 10.1073/pnas.68.6.1331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lim L., Canellakis E. S. Adenine-rich polymer associated with rabbit reticulocyte messenger RNA. Nature. 1970 Aug 15;227(5259):710–712. doi: 10.1038/227710a0. [DOI] [PubMed] [Google Scholar]
  18. Lodish H. F. Species specificity of polypeptide chain initiation. Nature. 1969 Nov 29;224(5222):867–870. doi: 10.1038/224867a0. [DOI] [PubMed] [Google Scholar]
  19. Smith A. E., Marcker K. A. Cytoplasmic methionine transfer RNAs from eukaryotes. Nature. 1970 May 16;226(5246):607–610. doi: 10.1038/226607a0. [DOI] [PubMed] [Google Scholar]
  20. Steitz J. A. Polypeptide chain initiation: nucleotide sequences of the three ribosomal binding sites in bacteriophage R17 RNA. Nature. 1969 Dec 6;224(5223):957–964. doi: 10.1038/224957a0. [DOI] [PubMed] [Google Scholar]
  21. Young R. J., Content J. 5'-terminus of influenza virus RNA. Nat New Biol. 1971 Mar 31;230(13):140–142. doi: 10.1038/newbio230140a0. [DOI] [PubMed] [Google Scholar]

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