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PMID: 22869748 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Diphthamide modification on eukaryotic elongation factor 2 is needed to assure fidelity of mRNA translation and mouse development.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 109 ·No. 34 ·2012-08-21 ·Pages 13817-22

Liu S, Bachran C, Gupta P, Miller-Randolph S, Wang H, Crown D, Zhang Y, Wein AN, Singh R, Fattah R, Leppla SH

Abstract

To study the role of the diphthamide modification on eukaryotic elongation factor 2 (eEF2), we generated an eEF2 Gly(717)Arg mutant mouse, in which the first step of diphthamide biosynthesis is prevented. Interestingly, the Gly(717)-to-Arg mutation partially compensates the eEF2 functional loss resulting from diphthamide deficiency, possibly because the added +1 charge compensates for the loss of the +1 charge on diphthamide. Therefore, in contrast to mouse embryonic fibroblasts (MEFs) from OVCA1(-/-) mice, eEF2(G717R/G717R) MEFs retain full activity in polypeptide elongation and have normal growth rates. Furthermore, eEF2(G717R/G717R) mice showed milder phenotypes than OVCA1(-/-) mice (which are 100% embryonic lethal) and a small fraction survived to adulthood without obvious abnormalities. Moreover, eEF2(G717R/G717R)/OVCA1(-/-) double mutant mice displayed the milder phenotypes of the eEF2(G717R/G717R) mice, suggesting that the embryonic lethality of OVCA1(-/-) mice is due to diphthamide deficiency. We confirmed that the diphthamide modification is essential for eEF2 to prevent -1 frameshifting during translation and show that the Gly(717)-to-Arg mutation cannot rescue this defect.

MeSH Terms
Adenosine Diphosphate/chemistry Animals Biotin/chemistry CHO Cells Cricetinae Fibroblasts/cytology Gene Deletion Histidine/analogs & derivatives,pharmacology Mice Mice, Transgenic Minor Histocompatibility Antigens Peptide Elongation Factor 2/metabolism Peptides/chemistry Phenotype Protein Biosynthesis Tumor Suppressor Proteins/genetics
Chemicals
Dph1 protein, mouse Minor Histocompatibility Antigens Peptide Elongation Factor 2 Peptides Tumor Suppressor Proteins Histidine Adenosine Diphosphate Biotin diphthamide
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Liu Shihui
Microbial Pathogenesis Section, Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. [email protected]
Bachran Christopher
Gupta Pradeep
Miller-Randolph Sharmina
Wang Hailun
Crown Devorah
Zhang Yi
Wein Alexander N
Singh Rajat
Fattah Rasem
Leppla Stephen H
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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
1091-6490
DOI
10.1073/pnas.1206933109
Email
[email protected]
Published
2012-08-21
Epub
2012-00-06
Pages
13817-22
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC3427129
Subset
IM
Grants
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1073/pnas.1206933109 PMC Full Text View journal details
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