Purification and characterization of mRNA cap-binding protein from Drosophila melanogaster embryos.

Molecular and Cellular Biology
F G Maroto, J M Sierra

Abstract

A protein with specific affinity for the mRNA cap structure was purified both from the postribosomal supernatant and from the ribosomal high-salt wash of Drosophila melanogaster embryos by m7GTP-Sepharose chromatography. This protein had an apparent molecular mass of 35 kilodaltons (kDa) in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a size very different from those of the cap-binding proteins that have been characterized thus far. Drosophila 35-kDa cap-binding protein (CBP) could also be isolated from the ribosomal high-salt wash as part of a salt-stable protein complex consisting of polypeptides of 35, 72, and 140 to 180 kDa. Polyclonal antibodies against Drosophila 35-kDa CBP neither reacted with eucaryotic initiation factor 4E from rabbit reticulocytes nor affected mRNA translation in a rabbit reticulocyte cell-free system. However, in a cell-free system from Drosophila embryos, mRNA translation was specifically inhibited by these antibodies. The requirement of 35-kDa CBP for mRNA translation in Drosophila was diminished under ionic conditions in which the importance of mRNA cap structure recognition was reduced. Despite the structural differences between Drosophila 35-kDa CBP and mammalian initiation factor ...Continue Reading

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Citations

Aug 2, 2008·Biochemical and Biophysical Research Communications·Raquel Reyes, José M Izquierdo
Jun 6, 2012·Comparative and Functional Genomics·Greco HernándezArmen Parsyan
Apr 18, 2012·Biochimica Et Biophysica Acta·Paola V FerreroRolando V Rivera-Pomar
Apr 5, 2005·Mechanisms of Development·Greco HernándezRolando Rivera-Pomar
Jun 1, 2005·Mechanisms of Development·Greco Hernández, Paula Vazquez-Pianzola
Jun 30, 2012·Comparative and Functional Genomics·Gritta TettweilerGreco Hernández
Jun 24, 2006·Cell Death and Differentiation·P Vazquez-PianzolaR Rivera-Pomar
May 1, 1994·Molecular Biology Reports·J M Sierra, J M Zapata
Apr 26, 1995·Biochimica Et Biophysica Acta·G Hernández, J M Sierra

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