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PMID: 3855554 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Isolation and characterization of factors from wheat germ that exhibit eukaryotic initiation factor 4B activity and overcome 7-methylguanosine 5'-triphosphate inhibition of polypeptide synthesis.

Lax S, Fritz W, Browning K, Ravel J

Abstract

Three highly purified preparations (preparations I, II-1, and II-2) have been obtained from wheat germ and shown to support in vitro polypeptide synthesis directed by capped or uncapped mRNAs in a eukaryotic initiation factor 4B (eIF-4B)-deficient system. The three preparations differ, however, in polypeptide composition and in the ability to overcome the inhibitory effect of 7-methylguanosine 5'-triphosphate (m7GTP) on in vitro polypeptide synthesis. Preparation I contains two polypeptides (Mr = 80,000 and 28,000), which are present in a 1:1 molar ratio and are associated in a high molecular weight complex. Preparation II-1 contains two major polypeptides (Mr = 220,000 and 26,000) and preparation II-2 also contains two major polypeptides (Mr = 110,000 and 26,000). Preparations II-1 and II-2 are high molecular weight complexes; neither contains detectable amounts of a Mr 80,000 or a Mr 50,000 component. Preparations II-1 and II-2 both overcome m7GTP inhibition, whereas preparation I does not. These findings raise several questions with regard to the identity of eIF-4B and its relationship to cap recognition factors.

MeSH Terms
Chromatography, Ion Exchange Eukaryotic Initiation Factors Molecular Weight Peptide Biosynthesis Peptide Initiation Factors/isolation & purification RNA Cap Analogs/pharmacology RNA Caps/pharmacology RNA, Messenger/metabolism Triticum/analysis
Chemicals
Eukaryotic Initiation Factors Peptide Initiation Factors RNA Cap Analogs RNA Caps RNA, Messenger eIF-4B 7-methylguanosine triphosphate
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lax S
Fritz W
Browning K
Ravel J
References (21)
21 references, click to expand
  1. Protein synthesis initiation factors from human HeLa cells and rabbit reticulocytes are similar: comparison of protein structure, activities, and immunochemical properties.
    Biochemistry. 1982 Aug 31;21(18):4202-6 PMID: 6181805
  2. Differential stimulation of capped mRNA translation in vitro by cap binding protein.
    Nature. 1980 May 29;285(5763):331-3 PMID: 6246452
  3. New initiation factor activity required for globin mRNA translation.
    J Biol Chem. 1983 May 10;258(9):5804-10 PMID: 6853548
  4. Involvement of eukaryotic initiation factor 4A in the cap recognition process.
    J Biol Chem. 1983 Sep 25;258(18):11398-403 PMID: 6604056
  5. Purification of the messenger RNA cap-binding protein using a new affinity medium.
    Biochemistry. 1984 Jan 17;23(2):177-81 PMID: 6696877
  6. In vitro translation of STNV-RNA.
    Methods Enzymol. 1974;30:754-61 PMID: 4604593
  7. Characterization of initiation factor eIF-3 from wheat germ.
    J Biol Chem. 1981 Feb 25;256(4):1582-6 PMID: 6906354
  8. Two forms of purified m7G-cap binding protein with different effects on capped mRNA translation in extracts of uninfected and poliovirus-infected HeLa cells.
    J Biol Chem. 1981 Aug 10;256(15):7691-4 PMID: 6267020
  9. A polypeptide which reverses cap analogue inhibition of cell-free protein synthesis. Purification and binding to capped oligonucleotides.
    J Biol Chem. 1982 Apr 25;257(8):4056-62 PMID: 7068625
  10. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  11. Functional characterization of eukaryotic mRNA cap binding protein complex: effects on translation of capped and naturally uncapped RNAs.
    Biochemistry. 1984 May 22;23(11):2456-62 PMID: 6089873
  12. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  13. Purification and characterization of initiation factors IF-E4 and IF-E6 from rabbit reticulocytes.
    J Biol Chem. 1977 Aug 25;252(16):5798-803 PMID: 885883
  14. Initiation of mammalian protein synthesis. I. Purification and characterization of seven initiation factors.
    J Mol Biol. 1977 Nov;116(4):727-53 PMID: 592398
  15. Partial purification of the factors required for the initiation of protein synthesis in wheat germ.
    J Biol Chem. 1979 Jan 10;254(1):143-8 PMID: 758313
  16. A polypeptide in eukaryotic initiation factors that crosslinks specifically to the 5'-terminal cap in mRNA.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4843-7 PMID: 217002
  17. Purification of seven protein synthesis initiation factors from Krebs II ascites cells.
    Biochim Biophys Acta. 1979 Feb 27;561(2):484-90 PMID: 427168
  18. Purification of protein synthesis initiation factors from rabbit reticulocytes.
    Methods Enzymol. 1979;60:101-8 PMID: 459891
  19. Isolation and purification of protein synthesis initiation factors from wheat germ.
    Methods Enzymol. 1979;60:193-204 PMID: 459899
  20. Eukaryotic mRNA cap binding protein: purification by affinity chromatography on sepharose-coupled m7GDP.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4345-9 PMID: 291969
  21. Fractionation and partial characterization of the protein synthesis system of wheat germ. I. Resolution of two elongation factors and five initiation factors.
    J Biol Chem. 1983 Jan 25;258(2):859-65 PMID: 6549754
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
1985-01-00
Pages
330-3
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC397031
Subset
IM
Grants
NIGMS NIH HHS · GM18775 · United States
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