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
-
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
-
Differential stimulation of capped mRNA translation in vitro by cap binding protein.
Nature. 1980 May 29;285(5763):331-3
PMID: 6246452
-
New initiation factor activity required for globin mRNA translation.
J Biol Chem. 1983 May 10;258(9):5804-10
PMID: 6853548
-
Involvement of eukaryotic initiation factor 4A in the cap recognition process.
J Biol Chem. 1983 Sep 25;258(18):11398-403
PMID: 6604056
-
Purification of the messenger RNA cap-binding protein using a new affinity medium.
Biochemistry. 1984 Jan 17;23(2):177-81
PMID: 6696877
-
In vitro translation of STNV-RNA.
Methods Enzymol. 1974;30:754-61
PMID: 4604593
-
Characterization of initiation factor eIF-3 from wheat germ.
J Biol Chem. 1981 Feb 25;256(4):1582-6
PMID: 6906354
-
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
-
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
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
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
-
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
-
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
-
Initiation of mammalian protein synthesis. I. Purification and characterization of seven initiation factors.
J Mol Biol. 1977 Nov;116(4):727-53
PMID: 592398
-
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
-
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
-
Purification of seven protein synthesis initiation factors from Krebs II ascites cells.
Biochim Biophys Acta. 1979 Feb 27;561(2):484-90
PMID: 427168
-
Purification of protein synthesis initiation factors from rabbit reticulocytes.
Methods Enzymol. 1979;60:101-8
PMID: 459891
-
Isolation and purification of protein synthesis initiation factors from wheat germ.
Methods Enzymol. 1979;60:193-204
PMID: 459899
-
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
-
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