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  Identification and characterization of the expression of the translation initiation factor 4A (eIF4A) from Drosophila melanogaster

Hernandez, G., Lalioti, V., Vandekerckhove, J., Sierra, J. M., & Santaren, J. F. (2004). Identification and characterization of the expression of the translation initiation factor 4A (eIF4A) from Drosophila melanogaster. Proteomics, 4: DOI 10.1002/pmic.2003300555, pp. 316-326. Retrieved from http://www3.interscience.wiley.com/cgi-bin/fulltext/104550696/PDFSTART.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-EE4C-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E547-2
Genre: Journal Article

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Hernandez, G.1, Author              
Lalioti, V., Author
Vandekerckhove, J., Author
Sierra, J. M., Author
Santaren, J. F., Author
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

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Free keywords: Drosophila; elF4A; Protein synthesis; Two-dimensional polyacrylamide gel electrophoresis
 Abstract: We have identified the initiation factor 4A (eIF4A) in a two-dimensional protein database of Drosophila wing imaginal discs. eIF4A, a member of the DEAD-box family of RNA helicases, forms the active eIF4F complex that in the presence of eIF4B and eIF4H unwinds the secondary structure of the 5’-UTR of mRNAs during translational initiation. Two-dimensional gel electrophoresis and microsequencing allowed us to purify eIF4A, and generate specific polyclonal antibodies. A combination of immunoblotting and labelling with [35S]methionine 1 [35S]cysteine revealed the existence of a single eIF4A isoform encoded by a previously reported gene that maps to chromosome 2L at 26A7-9. Expression of this gene yields two mRNA species, generated by alternative splicing in the 3’-untranslated region. The two mRNAs contain the same open reading frame and produce the identical eIF4A protein. No expression was detected of the eIF4A-related gene CG7483. We detected eIF4A protein expression in the wing imaginal discs of several Drosophila species, and in haltere, leg 1, leg 2, leg 3, and eyeantenna imaginal discs of D. melanogaster. Examination of eIF4A in tumor suppressor mutants showed significantly increased (>50%) expression in the wing imaginal discs of these larvae. We observed ubiquitous expression of eIF4A mRNA and protein during Drosophila embryogenesis. Yeast two-hybrid analysis demonstrated the in vivo interaction of Drosophila eIF4G with the N-terminal third of eIF4A.

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 Dates: 2004-02-162004-02-01
 Publication Status: Published in print
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 Rev. Method: Peer
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Title: Proteomics
Source Genre: Journal
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Pages: - Volume / Issue: 4 Sequence Number: DOI 10.1002/pmic.2003300555 Start / End Page: 316 - 326 Identifier: -