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ID: 63298.0, MPI für biophysikalische Chemie / Molekulare Biologie (Dr. Thomas M. Jovin)
Identification and characterization of the expression of the translation initiation factor 4A (eIF4A) from Drosophila melanogaster
Authors:Hernandez, G.; Lalioti, V.; Vandekerckhove, J.; Sierra, J. M.; Santaren, J. F.
Date of Publication (YYYY-MM-DD):2004-02-01
Title of Journal:Proteomics
Start Page:316
End Page:326
Sequence Number of Article:DOI 10.1002/pmic.2003300555
Review Status:Peer-review
Audience:Not Specified
Abstract / Description: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.
Free Keywords:Drosophila; elF4A; Protein synthesis; Two-dimensional polyacrylamide gel electrophoresis
Last Change of the Resource (YYYY-MM-DD):2004-02-16
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für biophysikalische Chemie/Abt. Thomas Jovin / 060
External Affiliations:Centro de Biologia Molecular "Severo Ochoa", Universidad Autónoma de Madrid and C.S.I.C., Madsrin, Spain / Department of Medical Protein Research, Flanders Interuniversity Insititue of Biotechnology, Department of Biochemsitry, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium
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