Influence of DPH1 and DPH5 Protein Variants on the Synthesis of Diphthamide, the Target of ADPRibosylating Toxins
Abstract
:1. Introduction
2. Results
2.1. Transient Expression of DPH1 and DPH5 cDNA in DPH Deficient MCF7 Cells
2.2. Plasmid Encoded DPH Proteins Restore Diphthamide Synthesis and Toxin Sensitivity in DPH1 or DPH5 Deficient Cells
2.3. DPH1 and DPH5 Variants
2.4. Activity of DPH1 and DPH5 Protein Variants
3. Discussion
4. Materials and Methods
4.1. MCF7 Variants with Inactivated DPH Genes
4.2. Expression of DPH Proteins
4.3. ADP-Ribosylation of eEF2
4.4. Cytotoxicity Assays
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Protein | Variant | Type | Source | Expression * | Phenotype |
---|---|---|---|---|---|
DPH1 | - | reference sequence (wt) | Uniprot Q9BZG8-1 | +++ | functional |
isotype 2 | N-term deletion (1-80) | Uniprot Q9BZG8-2 | ++ | loss of function | |
isotype 3 | N-term deletion (1-140) | Uniprot Q9BZG8-3 | +++ | loss of function | |
R27W | missense | COSM 178273 | + | functional | |
S56F | missense | COSM 389195 | + | functional | |
S221P | missense | COSM 1381407* | ++ | reduced function | |
L96 * | frameshift-stop | COSM 190903 | ? | loss of function | |
R312 * | nonsense-stop | COSM 137079 | ? | reduced function | |
DPH5 | - | reference sequence (wt) | Uniprot Q9H2P9-1 | +-+++ | functional |
del A212 | single aa deletion isoform | Uniprot Q9H2P9-6 | ++ | functional | |
D57G | missense | COSM 893022 | +++ | functional | |
G87R | missense | dbSNP rs376902046 | ++ | functional | |
S123C | missense | COSM 4792156 | +++ | functional | |
Q170H | missense | COSM 674720 | + | functional | |
E60 * | nonsense-stop | COSM 260571 | ? | loss of function | |
W136 * | frameshift-stop | COSM 5002373 | ? | loss of function | |
R207 * | nonsense-stop | COSM 3417782 | ? | loss of function |
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Mayer, K.; Schröder, A.; Schnitger, J.; Stahl, S.; Brinkmann, U. Influence of DPH1 and DPH5 Protein Variants on the Synthesis of Diphthamide, the Target of ADPRibosylating Toxins. Toxins 2017, 9, 78. https://doi.org/10.3390/toxins9030078
Mayer K, Schröder A, Schnitger J, Stahl S, Brinkmann U. Influence of DPH1 and DPH5 Protein Variants on the Synthesis of Diphthamide, the Target of ADPRibosylating Toxins. Toxins. 2017; 9(3):78. https://doi.org/10.3390/toxins9030078
Chicago/Turabian StyleMayer, Klaus, Anna Schröder, Jerome Schnitger, Sebastian Stahl, and Ulrich Brinkmann. 2017. "Influence of DPH1 and DPH5 Protein Variants on the Synthesis of Diphthamide, the Target of ADPRibosylating Toxins" Toxins 9, no. 3: 78. https://doi.org/10.3390/toxins9030078