Characterization of the endogenous ADP‐ribosylation of wild‐type and mutant elongation factor 2 in eukaryotic cells

Fendrick, James L.; Iglewski, Wallace J.; Moehring, Joan M.; Moehring, Thomas J.

doi:10.1111/j.1432-1033.1992.tb16748.x

Cited by 34 publications

(25 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There have been some reports suggesting the existence of cellular ADP-transribosylase: in polyoma virus-transformed baby hamster kidney cells, the same peptide that can be ADP-ribosylated by diphtheria toxin has also been shown to be modiˆed, and higher ribosylation was observed at 2z than 10z serum in the culture medium. 72) These results suggest that ADP-ribosylation would control EF-2 activity under nutritional deprivation. However, the enzymes that transfer or remove the ADP moiety from EF- 2 have not yet been isolated.…”

Section: Diseases and Elongation Factorsmentioning

confidence: 78%

Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin Bundling to Zinc Finger Protein R1-Associated Nuclear Localization

Ejiri

2002

Bioscience, Biotechnology, and Biochemistry

190

167

View full text Add to dashboard Cite

Section: Diseases and Elongation Factorsmentioning

confidence: 78%

Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin Bundling to Zinc Finger Protein R1-Associated Nuclear Localization

Ejiri

2002

Bioscience, Biotechnology, and Biochemistry

190

167

View full text Add to dashboard Cite

“…The mAf1521 not only binds the G protein ␤ subunit that we have shown to be modified on arginine (17) but also GDH that is modified on cysteine (18), eEF2 on diphthamide (7,31), and GRP78/BiP on an unidentified amino acid, clearly indicating that the recognition of the ADP-ribose is independent of the modified amino acid. This highlights the potential of our methodology and its relevance for the definition of the role of the ADP-ribosylation reaction.…”

Section: Purification and Identification Of Substrates Of Intracellulmentioning

confidence: 99%

Combining affinity purification by ADP-ribose-binding macro domains with mass spectrometry to define the mammalian ADP-ribosyl proteome

Dani

Stilla

Marchegiani

et al. 2009

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

104

View full text Add to dashboard Cite

Mono-ADP-ribosylation is a reversible posttranslational modification that modulates the function of target proteins. The enzymes that catalyze this reaction in mammalian cells are either bacterial pathogenic toxins or endogenous cellular ADP-ribosyltransferases. For the latter, both the enzymes and their targets have largely remained elusive, mainly due to the lack of specific techniques to study this reaction. The recent discovery of the macro domain, a protein module that interacts selectively with ADP-ribose, prompted us to investigate whether this interaction can be extended to the identification of ADP-ribosylated proteins. Here, we report that macro domains can indeed be used as selective baits for high-affinity purification of mono-ADP-ribosylated proteins, which can then be identified by mass spectrometry. Using this approach, we have identified a series of cellular targets of ADP-ribosylation reactions catalyzed by cellular ADP-ribosyltransferases and toxins. These proteins include most of the known targets of ADP-ribosylation, indicating the validity of this method, and a large number of other proteins, which now need to be individually validated. This represents an important step toward the discovery of new ADP-ribosyltransferase targets and an understanding of the physiological role and the pharmacological potential of this protein modification.ADP-ribosylation ͉ ADP-ribosyltransferase ͉ NAD ͉ toxin ͉ posttranslational modification

show abstract

“…One cDNA plasmid (p17 in ref. 1) was particularly interesting because the phenotype it produced could not be attributed to known mechanisms of PE and DT resistance (2,3). In fact, when cells transfected with this plasmid were exposed to toxin, modification of elongation factor EF2 and inhibition of protein synthesis was comparable to toxin-sensitive controls, yet the cells did not die.…”

mentioning

confidence: 99%

Cloning and characterization of a cellular apoptosis susceptibility gene, the human homologue to the yeast chromosome segregation gene CSE1.

Brinkmann

Gallo

et al. 1995

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

162

148

View full text Add to dashboard Cite

We recently isolated human cDNA fragments that render MCF-7 breast cancer cells resistant to cell death caused by Pseudomonas exotoxin, Pseudomonas exotoxinderived immunotoxins, diphtheria toxin, and tumor necrosis factor. We report here that one of these fragments is We have used expression cloning to isolate cDNAs containing plasmids that cause MCF-7 breast cancer cells to become resistant to immunotoxins (1). These plasmids also render the cells resistant to native Pseudomonas exotoxin (PE) and diphtheria toxin (DT); thus the resistance is due to the action of the toxin moiety of immunotoxins and not due to reduced expression of the antigen to which the immunotoxin is directed. One cDNA plasmid (p17 in ref. 1) was particularly interesting because the phenotype it produced could not be attributed to known mechanisms of PE and DT resistance (2, 3). In fact, when cells transfected with this plasmid were exposed to toxin, modification of elongation factor EF2 and inhibition of protein synthesis was comparable to toxin-sensitive controls, yet the cells did not die. Thus, this cDNA appears to be involved in a mechanism affecting the sensitivity of cells to toxin after the primary action of the toxin, inhibition of protein synthesis, has occurred. The observation that PE and DT can induce apoptosis (4-6) and the observation that this plasmid also confers resistance to tumor necrosis factor (TNF) a and ,B indicate that this cDNA could mediate resistance by interference with apoptosis (unpublished results). Here we describe that the isolated resistance plasmid that mediates toxin as well as TNF resistance to MCF-7 cells contains an antisense cDNA fragment homologous to the yeast CSE1 gene (7). We report the sequence of the human CSE1 homologue and present evidence that this gene may play a role in cell proliferation.t MATERIALS AND METHODSExpression Cloning and Immunotoxin Selection of cDNA Plasmids. Human cDNAs that confer resistance to the immunotoxin B3(Fv)-PE38KDEL were isolated by expression cloning and immunotoxin selection from a cDNA library in pCDM8 (HeLa cDNA expressed from a cytomegalovirus promoter followed by simian virus 40 poly(A) as described (1). B3(Fv)-PE38KDEL is a fusion protein composed of a truncated form of PE and the Fv region of monoclonal antibody B3 that binds to a carbohydrate present on many carcinomas and cancer cell lines-e.g., on MCF-7 cells-and kills such cells (8). pCDM/HE17 (p17 in ref. 1) is a plasmid that confers resistance not only to immunotoxin, PE, and DT but also to TNF.

show abstract

Characterization of the endogenous ADP‐ribosylation of wild‐type and mutant elongation factor 2 in eukaryotic cells

Cited by 34 publications

References 29 publications

Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin Bundling to Zinc Finger Protein R1-Associated Nuclear Localization

Moonlighting Functions of Polypeptide Elongation Factor 1: From Actin Bundling to Zinc Finger Protein R1-Associated Nuclear Localization

Combining affinity purification by ADP-ribose-binding macro domains with mass spectrometry to define the mammalian ADP-ribosyl proteome

Cloning and characterization of a cellular apoptosis susceptibility gene, the human homologue to the yeast chromosome segregation gene CSE1.

Contact Info

Product

Resources

About