Primary structure of elongation factor 1γ from <i>Artemia</i>

Maessen, G.D.F.; Amons, Reinout; Zeelen, J.P.; Möller, W.

doi:10.1016/0014-5793(87)80532-x

Cited by 36 publications

(15 citation statements)

References 21 publications

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“…These protein bands (X and Y) were then blotted onto a poly(vinylidene difluoride) membrane (9) and subjected to microsequence analyses (10) followed by a homology search from a protein sequence database with the BLAST program. As shown in Table 1, the three peptide fragments derived from protein band X were 72-88% identical in amino acid sequence to the protein synthesis elongation factor EF-1␥ of brine shrimp, Artemia salina (13). On the other hand, the two peptide fragments derived from protein band Y bear 82 and 88% amino acid sequence identity with the protein synthesis elongation factor EF-1␤ of silkworm (14).…”

Section: Characterization Of Cellular Factor(s)mentioning

confidence: 92%

RNA polymerase of vesicular stomatitis virus specifically associates with translation elongation factor-1 αβγ for its activity

Das

Mathur

Gupta

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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An RNA-dependent RNA polymerase is packaged within the virions of purified vesicular stomatitis virus, a nonsegmented negative-strand RNA virus, which carries out transcription of the genome RNA into mRNAs both in vitro and in vivo. The RNA polymerase is composed of two virally encoded polypeptides: a large protein L (240 kDa) and a phosphoprotein P (29 kDa). Recently, we obtained biologically active L protein from insect cells following infection by a recombinant baculovirus expressing L gene. During purification of the L protein from Sf21 cells, we obtained in addition to an active L fraction an inactive fraction that required uninfected insect cell extract to restore its activity. The cellular factors have now been purified, characterized, and shown to be ␤ and ␥ subunits of the protein synthesis elongation factor EF-1. We also demonstrate that the ␣ subunit of EF-1 remains tightly bound to the L protein in the inactive fraction and ␤␥ subunits associate with the L(␣) complex. Further purification of L(␣) from the inactive fraction revealed that the complex is partially active and is significantly stimulated by the addition of ␤␥ subunits purified from Sf21 cells. A putative inhibitor(s) appears to co-elute in the inactive fraction that blocked the L(␣) activity. The purified virions also package all three subunits of EF-1. These findings have a striking similarity with Q␤ RNA phage, which also associates with the bacterial homologue of EF-1 for its replicase function, implicating a possible evolutionary relationship between these host proteins and the RNA-dependent RNA polymerase of RNA viruses.Vesicular stomatitis virus (VSV), a prototype of nonsegmented negative-strand RNA viruses, has long been a paradigm for studying gene expression of this class of RNA viruses that infect vertebrates, invertebrates, and plants (1). Some of the most common human pathogens that belong to this category are rabies, measles, mumps, and human parainfluenza. A hallmark of all negative strand RNA viruses is the obligate packaging of an RNA-dependent RNA polymerase within the mature virions (2) that transcribes the genome RNA into mRNAs both in vitro and in vivo (3). For VSV, the virion-associated RNA polymerase is generally thought to consist of two virally encoded protein subunits, L (240 kDa) and P (29 kDa), which remain tightly complexed within the virion (3). Studies on the structure and function of VSV RNA polymerase have been greatly aided by the ability to isolate the polymerase subunits from the virions in a relatively pure form (4, 5). Active reconstitution of transcription is achieved by mixing the genome RNA enwrapped with the nucleocapsid protein (N) (referred to as N-RNA template) and purified L and P proteins (4, 5). From a large body of evidence, it appears that the L protein possesses the catalytic activity for RNA synthesis and the P protein is a transcription factor essential for L function (3); no cellular protein(s) has so far been shown to be required for the RNA polymerase activity. Only recently, ...

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Section: Characterization Of Cellular Factor(s)mentioning

confidence: 92%

RNA polymerase of vesicular stomatitis virus specifically associates with translation elongation factor-1 αβγ for its activity

Das

Mathur

Gupta

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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“…The four largest peptides (19,23,26 and 27 kDa) arose within a few minutes upon cleavage of EF-1y in this labile region (Fig. 4).…”

Section: Proteolytic Digestion Of Ef-ipymentioning

confidence: 95%

Mapping the functional domains of the eukaryotic elongation factor 1βγ

Damme¹,

Amons²,

Janssen³

et al. 1991

European Journal of Biochemistry

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The functional domains of the eukaryotic elongation factor (EF) 1Py have been delineated with the use of limited proteolysis, protein microsequencing, gel electrophoresis under non-denaturing conditions and antibodies against EF-1P and EF-ly. By means of limited proteolysis, it was possible to obtain large fragments of EF-ID. In contrast to amino-terminal fragments, those derived from the carboxy-terminal part of EF-ID were still active in enhancing the guanine nucleotide exchange of GDP bound to EF-la. With the same technique of limited proteolysis, it was possible to isolate a trypsin-resistant core from EF-1 By containing polypeptide chain fragments derived from both subunits. A polyvalent antiserum against EF-1P and two monoclonal antibodies against EFl y were used to identify the protein fragments in this core. The monoclonal antibodies were shown to recognize different epitopes, one localized on the amino-terminal and another on the carboxy-terminal half of EF-I y. The antiserum against EF-1P and one of the monoclonal antibodies (mAb 36E5), which recognized the amino-terminal half of EF-ly, reacted with this trypsin-resistant core. We conclude that the amino-terminal halves of both EF-IB and EF-ly are firmly attached to each other, and that the carboxy-terminal part of EF-lP interacts with EF-la.Protein synthesis requires factors for proper translation [I]. In prokaryotes, correct binding of aminoacyl tRNA to translating ribosomes is mediated by elongation factor EFTu, a protein which is able to form a ternary complex with aminoacyl tRNA and GTP. Under simultaneous hydrolysis of GTP into GDP and Pi, the cognate aminoacyl tRNA becomes bound to the ribosome mRNA complex [2]. Replacement of GDP, in the EF-Tu . GDP complex, by GTP is enhanced by a separate factor, EF-Ts, the nucleotide-exchange factor [2]. The situation in eukaryotes is similar; here, however, EF-I a ( Z 50 kDa) replaces EF-Tu, whereas a complex between two polypeptide chains, EF-1fi and EF-ly, functions as the nucleotide-exchange factor [2]. The actual exchange activity, which resides on EF-lP (~2 6 kDa), is modulated by phosphorylation of its residue Ser89 [3]. The role of EF-ly (~4 6 kDa), a protein with strong hydrophobic properties, is not yet clear, although it possesses a stimulating effect on the exchange activity of EF-1P [4]. High-molecular-mass complexes between EF-la and EF-1Py have been described in many eukaryotic tissues [5 -111.In an attempt to unravel the requirements for nucleotideexchange activity and the involvement of the protein stretch around the phosphorylated residue Ser89 in EF-IP, we have isolated large, well-defined proteolytic fragments of this pro- Abbreviations. EF-la, EF-Ib, EF-Iy and EF-16, the a, b, y and 6 subunits of the eukaryotic protein synthesis elongation factor 1 ; EFTu, prokaryotic equivalent of the eukaryotic protein synthesis elongation factor 13; EF-Ts, prokaryotic equivalent of the eukaryotic protein synthesis elongation factor 1 f l y ; mAb, monoclonal antibody.Enzymes. Trypsin (EC 3.4.21.4); clo...

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“…from shrimp Artemia salina [9] as depicted in fig.3. Since EF-I~ has been described to form a complex in association with EF-17 [18], we have searched for homology between p47 polypeptides and EF-17 [10].…”

Section: Methodsmentioning

confidence: 99%

“…We report here using amino acid analysis of peptides, that the p30 and p47 proteins of the complex are respectively analogous to EF-I~ [9] and EF-17 [10] from Artemia salina.…”

Section: Introductionmentioning

confidence: 92%

A purified complex from Xenopus oocytes contains a p47 protein, an in vivo substrate of MPF, and a p30 protein respectively homologous to elongation factors EF‐1γ and EF‐1β

et al. 1989

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A high molecular mass complex isolated from Xenopus laevis oocytes contains three main proteins, respectively p30, p36 and p47. The p47 protein has been reported to be an in vivo substrate of the cell division control protein kinase p34 '~2. From polypeptide sequencing, we now show that the p30 and the p47 correspond to elongation factor EF-lfl and EF-ly.Furthermore, the p30 and p36 proteins were phosphorylated in vitro by casein kinase II.

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Primary structure of elongation factor 1γ from Artemia

Cited by 36 publications

References 21 publications

RNA polymerase of vesicular stomatitis virus specifically associates with translation elongation factor-1 αβγ for its activity

RNA polymerase of vesicular stomatitis virus specifically associates with translation elongation factor-1 αβγ for its activity

Mapping the functional domains of the eukaryotic elongation factor 1βγ

A purified complex from Xenopus oocytes contains a p47 protein, an in vivo substrate of MPF, and a p30 protein respectively homologous to elongation factors EF‐1γ and EF‐1β

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