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PMID: 6760129 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Protein-RNA crosslinking in Escherichia coli 30S ribosomal subunits. Identification of a 16S rRNA fragment crosslinked to protein S12 by the use of the chemical crosslinking reagent 1-ethyl-3-dimethyl-aminopropylcarbodiimide.

Nucleic acids research ·Vol. 10 ·No. 23 ·1982-12-11 ·Pages 7657-76

Chiaruttini C, Expert-Bezançon A, Hayes D, Ehresmann B

Abstract

1-ethyl-3-dimethyl aminopropylcarbodiimide (EDC) was used to cross-link 30S ribosomal proteins to 16S rRNA within the E. coli 3OS ribosomal subunit. Covalently linked complexes containing 30S proteins and 16S rRNA, isolated by sedimentation of dissociated crosslinked 30S subunits through SDS containing sucrose gradients, were digested with RNase T1, and the resulting oligonucleotide-protein complexes were fractionated on SDS containing polyacrylamide gels. Eluted complexes containing 30S proteins S9 and S12 linked to oligonucleotides were obtained in pure form. Oligonucleotide 5'terminal labelling was successful in the case of S12 containing but not of the S9 containing complex and led to identification of the S12 bound oligonucleotide as CAACUCG which is located at positions 1316-1322 in the 16S rRNA sequence. Protein S12 is crosslinked to the terminal G of this heptanucleotide.

MeSH Terms
Bacterial Proteins/genetics Base Sequence Carbodiimides/pharmacology Cross-Linking Reagents/pharmacology Escherichia coli/genetics Ethyldimethylaminopropyl Carbodiimide/pharmacology RNA, Ribosomal/genetics Ribosomal Proteins/genetics
Chemicals
Bacterial Proteins Carbodiimides Cross-Linking Reagents RNA, Ribosomal Ribosomal Proteins ribosomal protein L7-L12 Ethyldimethylaminopropyl Carbodiimide
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chiaruttini C
Expert-Bezançon A
Hayes D
Ehresmann B
References (40)
40 references, click to expand
  1. Advances in the chemistry of carbodiimides.
    Chem Rev. 1967 Apr;67(2):107-52 PMID: 4859920
  2. The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins.
    Biochemistry. 1969 Jul;8(7):2897-905 PMID: 4897206
  3. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  4. Luteinizing hormone derivatives with covalently-linked subunits.
    FEBS Lett. 1974 Aug 25;44(2):224-8 PMID: 4472843
  5. The use of a new series of cleavable protein-crosslinkers on the Escherichia coli ribosome.
    FEBS Lett. 1974 Nov 15;48(2):288-92 PMID: 4373293
  6. Specific cross-linking of proteins S7 and L4 to ribosomal RNA, by UV irradiation of Escherichia coli ribosomal subunits.
    Mol Gen Genet. 1975 Dec 9;141(4):343-55 PMID: 814400
  7. 30S ribosomal proteins associated with the 3'-terminus of 16S RNA.
    FEBS Lett. 1975 Oct 15;58(1):281-4 PMID: 1225593
  8. A strain of Escherichia coli which gives rise to mutations in a large number of ribosomal proteins.
    Mol Gen Genet. 1976 Dec 22;149(3):303-9 PMID: 799249
  9. Identification of neighbouring proteins in Escherichia coli 30 S ribosome subunits.
    J Mol Biol. 1977 Jun 5;112(4):603-29 PMID: 327079
  10. The use of formaldehyde in RNA-protein cross-linking studies with ribosomal subunits from Escherichia coli.
    Eur J Biochem. 1977 Jun 1;76(1):175-87 PMID: 407080
  11. Characterization of the regions from E. coli 16 S RNA covalently linked to ribosomal proteins S4 and S20 after ultraviolet irradiation.
    FEBS Lett. 1977 Jun 15;78(2):261-6 PMID: 885249
  12. Mapping adenines, guanines, and pyrimidines in RNA.
    Nucleic Acids Res. 1977 Aug;4(8):2527-38 PMID: 409999
  13. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film.
    FEBS Lett. 1977 Oct 15;82(2):314-6 PMID: 913604
  14. The use of nuclease P1 in sequence analysis of end group labeled RNA.
    Nucleic Acids Res. 1977 Dec;4(12):4091-108 PMID: 202925
  15. Sequence analysis of 5'[32P] labeled mRNA and tRNA using polyacrylamide gel electrophoresis.
    Nucleic Acids Res. 1978 Jan;5(1):37-56 PMID: 25417
  16. Chemical cross-linking of protein to RNA within intact ribosomal subunits from Escherichia coli.
    Mol Gen Genet. 1978 Apr 6;160(2):183-93 PMID: 349353
  17. The sequence of Escherichia coli ribosomal 16 S RNA determined by new rapid gel methods.
    FEBS Lett. 1978 Oct 1;94(1):152-6 PMID: 359355
  18. RNA-protein neighbourhoods of the ribosome obtained by crosslinking.
    Eur J Biochem. 1978 Sep 1;89(2):353-9 PMID: 361392
  19. The use of sym-triazine trichloride in RNA-protein cross-linking studies with Escherichia coli ribosomal subunits.
    Mol Gen Genet. 1979 Jan 5;168(1):81-6 PMID: 372742
  20. The structure of the RNA binding site of ribosomal proteins S8 and S15.
    J Biol Chem. 1979 May 25;254(10):3873-8 PMID: 108264
  21. Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem, GFR.
    Nucleic Acids Res. 1979;6(5):1775-90 PMID: 377231
  22. Binding sites for ribosomal proteins S8 and S15 in the 16 S RNA of Escherichia coli.
    Biochim Biophys Acta. 1979 Jul 26;563(2):422-31 PMID: 380655
  23. Identification of a cytidine-specific ribonuclease from chicken liver.
    J Biol Chem. 1980 Mar 10;255(5):2160-3 PMID: 6986389
  24. Synthesis and properties of N-acetyl-N'-(p-glyoxylylbenzoyl)cystamine, a new reagent for RNA-RNA and RNA-protein cross-linking.
    Eur J Biochem. 1980 Jan;103(2):365-75 PMID: 6988212
  25. RNA-protein crosslinking by reaction with a soluble carbodiimide.
    FEBS Lett. 1980 Sep 22;119(1):145-9 PMID: 6159231
  26. Photochemical cross-linking of initiation factor-3 to Escherichia coli 30 S ribosomal subunits.
    J Biol Chem. 1980 Nov 10;255(21):10526-31 PMID: 7000779
  27. Secondary structure model for bacterial 16S ribosomal RNA: phylogenetic, enzymatic and chemical evidence.
    Nucleic Acids Res. 1980 May 24;8(10):2275-93 PMID: 6159576
  28. Synthesis of a new reagent, ethyl 4-azidobenzoylaminoacetimidate, and its use for RNA-protein cross-linking within Escherichia coli ribosomal 30-S subunits.
    Eur J Biochem. 1980 Sep;110(2):485-92 PMID: 7002551
  29. An experimentally-derived model for the secondary structure of the 16S ribosomal RNA from Escherichia coli.
    Nucleic Acids Res. 1980 Jun 11;8(11):2377-95 PMID: 6160459
  30. Phy M: an RNase activity specific for U and A residues useful in RNA sequence analysis.
    Nucleic Acids Res. 1980 Jul 25;8(14):3133-42 PMID: 6160466
  31. Identification of a 16-S RNA fragment crosslinked to protein S1 within Escherichia coli ribosomal 30-S subunits by the use of a crosslinking reagent: ethyl 4-azidobenzoylaminoacetimidate.
    Eur J Biochem. 1981 Apr;115(3):479-84 PMID: 6786883
  32. IF-3 crosslinking to Escherichia coli ribosomal 30 S subunits by three different light-dependent procedures: identification of 30 S proteins crosslinked to IF-3--utilization of a new two-stage crosslinking reagent, p-nitrobenzylmaleimide.
    Arch Biochem Biophys. 1981 May;208(2):554-62 PMID: 7020604
  33. The use of azidoarylimidoesters in RNA-protein cross-linking studies with Escherichia coli ribosomes.
    J Mol Biol. 1980 Mar 5;137(3):301-4 PMID: 7021847
  34. Structural organization of the 16S ribosomal RNA from E. coli. Topography and secondary structure.
    Nucleic Acids Res. 1981 May 11;9(9):2153-72 PMID: 6170942
  35. Ribonucleic acid-protein crosslinking in Escherichia coli ribosomal 30S subunits by the use of two new heterobifunctional reagents: 4-azido-2,3,5,6-tetrafluoropyridine and 4-azido-3,5-dichloro-2,6-difluoropyridine.
    Biochem Biophys Res Commun. 1981 Aug 14;101(3):784-91 PMID: 6171277
  36. The subunit interface of the Escherichia coli ribosome. Identification of proteins at the interface between the 30 S and 50 S subunits by crosslinking with 2-iminothiolane.
    J Mol Biol. 1981 Jul 5;149(3):451-76 PMID: 7031269
  37. A general secondary-structure model for procaryotic and eucaryotic RNAs from the small ribosomal subunits.
    Eur J Biochem. 1981 Dec;120(3):487-95 PMID: 6174326
  38. A new method for sequencing DNA.
    Proc Natl Acad Sci U S A. 1977 Feb;74(2):560-4 PMID: 265521
  39. Positions of proteins S6, S11 and S15 in the 30 S ribosomal subunit of Escherichia coli.
    J Mol Biol. 1981 Dec 15;153(3):739-60 PMID: 7040690
  40. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY.
    Biochem J. 1963 Oct;89:114-23 PMID: 14097352
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.23.7657
Published
1982-12-11
Pages
7657-76
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327037
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/10.23.7657 PMC Full Text View journal details
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