Reovirus MRNA Can Be Covalently Crosslinked Via the 5' Cap to Proteins in Initiation Complexes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1977 Oct 1

PMID 270673

Citations 28

Authors

N Sonenberg

A J Shatkin

Affiliations

Soon will be listed here.

Abstract

Proteins that are located adjacent to the 5' end of mRNA in initiation complexes have been detected by chemical crosslinking. Reovirus mRNA containing radioactivity exclusively in the [3H]methyl-labeled "cap," m7G(5')ppp(5')-Gm, was oxidized with sodium periodate to convert the 2',3'-cis-diol of the 5'-terminal m7G to a reactive dialdehyde. Oxidized mRNA was incubated in cell-free protein-synthesizing systems derived from wheat germ or mammalian cells, and the resulting mRNA-ribosome initiation complexes were reduced with NaBH3CN. By this chemical procedure, putative Schiff bases between mRNA 5'termini and amino groups of neighboring proteins were stabilized by reduction, yielding covalently linked protein-RNA conjugates. Under conditions of ribosome binding, a limited number of polypeptides associated with the mRNA-ribosome complexes were crosslinked, suggesting that these proteins are positioned near and may interact with the 5' end of mRNA during initiation. This method should also be useful for studying the spatial relationships between molecules in other similar nucleoprotein complexes.

Citing Articles

Immunoprecipitation Methods to Isolate Messenger Ribonucleoprotein Complexes (mRNP).

Hayek H, Gross L, Alghoul F, Martin F, Eriani G, Allmang C Adv Exp Med Biol. 2024; 3234:1-15.

PMID: 38507196 DOI: 10.1007/978-3-031-52193-5_1.

Bi-directional ribosome scanning controls the stringency of start codon selection.

Gu Y, Mao Y, Jia L, Dong L, Qian S Nat Commun. 2021; 12(1):6604.

PMID: 34782646 PMC: 8593136. DOI: 10.1038/s41467-021-26923-3.

Identification of the mAm Methyltransferase PCIF1 Reveals the Location and Functions of mAm in the Transcriptome.

Boulias K, Toczydlowska-Socha D, Hawley B, Liberman N, Takashima K, Zaccara S Mol Cell. 2019; 75(3):631-643.e8.

PMID: 31279658 PMC: 6703822. DOI: 10.1016/j.molcel.2019.06.006.

MicroRNAs recruit eIF4E2 to repress translation of target mRNAs.

Chen S, Gao G Protein Cell. 2017; 8(10):750-761.

PMID: 28755203 PMC: 5636748. DOI: 10.1007/s13238-017-0444-0.

Nm-seq maps 2'-O-methylation sites in human mRNA with base precision.

Dai Q, Moshitch-Moshkovitz S, Han D, Kol N, Amariglio N, Rechavi G Nat Methods. 2017; 14(7):695-698.

PMID: 28504680 PMC: 5712428. DOI: 10.1038/nmeth.4294.

References

Furuichi Y, Shatkin A . 5'-termini of reovirus mRNA: ability of viral cores to form caps post-transcriptionally. Virology. 1977; 77(2):566-78. DOI: 10.1016/0042-6822(77)90482-2. View

Furuichi Y, LaFiandra A, Shatkin A . 5'-Terminal structure and mRNA stability. Nature. 1977; 266(5599):235-9. DOI: 10.1038/266235a0. View

Furuichi Y, Shatkin A . Differential synthesis of blocked and unblocked 5'-termini in reovirus mRNA: effect of pyrophosphate and pyrophosphatase. Proc Natl Acad Sci U S A. 1976; 73(10):3448-52. PMC: 431132. DOI: 10.1073/pnas.73.10.3448. View

Bretner M, Sierakowska H, Szczesna E, Filipowicz W, Shatkin A . Removal of 5'-terminal m7G from eukaryotic mRNAs by potato nucleotide pyrophosphatase and its effect on translation. Nucleic Acids Res. 1977; 4(9):3065-81. PMC: 342635. DOI: 10.1093/nar/4.9.3065. View

Benne R, Hershey J . Purification and characterization of initiation factor IF-E3 from rabbit reticulocytes. Proc Natl Acad Sci U S A. 1976; 73(9):3005-9. PMC: 430909. DOI: 10.1073/pnas.73.9.3005. View

Shatkin A . Capping of eucaryotic mRNAs. Cell. 1976; 9(4 PT 2):645-53. DOI: 10.1016/0092-8674(76)90128-8. View

Muthukrishnan S, Morgan M, Banerjee A, Shatkin A . Influence of 5'-terminal m7G and 2'--O-methylated residues on messenger ribonucleic acid binding to ribosomes. Biochemistry. 1976; 15(26):5761-8. DOI: 10.1021/bi00671a012. View

SCHREIER M, Staehelin T . Initiation of mammalian protein synthesis: the importance of ribosome and initiation factor quality for the efficiency of in vitro systems. J Mol Biol. 1973; 73(3):329-49. DOI: 10.1016/0022-2836(73)90346-x. View

Crystal R, Elson N, Anderson W . Initiation of globin synthesis: assays. Methods Enzymol. 1974; 30:101-27. DOI: 10.1016/0076-6879(74)30014-6. View

10.

Eggen K, Shatkin A . In vitro translation of cardiovirus ribonucleic acid by mammalian cell-free extracts. J Virol. 1972; 9(4):636-45. PMC: 356348. DOI: 10.1128/JVI.9.4.636-645.1972. View

11.

HOWARD G, Traugh J, Croser E, Traut R . Ribosomal proteins from rabbit reticulocytes: number and molecular weights of proteins from ribosomal subunits. J Mol Biol. 1975; 93(3):391-404. DOI: 10.1016/0022-2836(75)90285-5. View

12.

Sundkvist I, Staehelin T . Structure and function of free 40 S ribosome subunits: Characterization of initiation factors. J Mol Biol. 1975; 99(3):401-18. DOI: 10.1016/s0022-2836(75)80135-5. View

13.

Safer B, Adams S, Kemper W, Berry K, Lloyd M, Merrick W . Purification and characterization of two initiation factors required for maximal activity of a highly fractionated globin mRNA translation system. Proc Natl Acad Sci U S A. 1976; 73(8):2584-8. PMC: 430692. DOI: 10.1073/pnas.73.8.2584. View

14.

Canaani D, Revel M, Groner Y . Translational discrimination of 'capped' and 'non-capped' mRNAS: inhibition of a series of chemical analogs of m7GpppX. FEBS Lett. 1976; 64(2):326-31. DOI: 10.1016/0014-5793(76)80321-3. View

15.

Legon S, Brayley A, Hunt T, Jackson R . The effect of cyclic AMP and related compounds on the control of protein synthesis in reticulocyte lysates. Biochem Biophys Res Commun. 1974; 56(3):745-52. DOI: 10.1016/0006-291x(74)90668-8. View

16.

Both G, Furuichi Y, Muthukrishnan S, Shatkin A . Effect of 5'-terminal structure and base composition on polyribonucleotide binding to ribosomes. J Mol Biol. 1976; 104(3):637-58. DOI: 10.1016/0022-2836(76)90126-1. View

17.

Both G, Furuichi Y, Muthukrishnan S, Shatkin A . Ribosome binding to reovirus mRNA in protein synthesis requires 5' terminal 7-methylguanosine. Cell. 1975; 6(2):185-95. DOI: 10.1016/0092-8674(75)90009-4. View

18.

Muthukrishnan S, Both G, Furuichi Y, Shatkin A . 5'-Terminal 7-methylguanosine in eukaryotic mRNA is required for translation. Nature. 1975; 255(5503):33-7. DOI: 10.1038/255033a0. View

19.

Both G, Banerjee A, Shatkin A . Methylation-dependent translation of viral messenger RNAs in vitro. Proc Natl Acad Sci U S A. 1975; 72(3):1189-93. PMC: 432492. DOI: 10.1073/pnas.72.3.1189. View

20.

Laemmli U, Favre M . Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973; 80(4):575-99. DOI: 10.1016/0022-2836(73)90198-8. View