Mapping the cleavage site in protein synthesis initiation factor eIF-4 gamma of the 2A proteases from human Coxsackievirus and rhinovirus
| Autor: | Hans Dieter Liebig, Fang Yang, Barry J. Lamphear, Hannes Klump, Robert E. Rhoads, Riqiang Yan, Tim Skern, Debra A. Waters, Ernst Kuechler |
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| Rok vydání: | 1993 |
| Předmět: |
Messenger RNA
Proteases Protease Edman degradation Protein subunit medicine.medical_treatment macromolecular substances Cell Biology Biology musculoskeletal system Cleavage (embryo) environment and public health Biochemistry Molecular biology health occupations Protein biosynthesis medicine Initiation factor heterocyclic compounds Molecular Biology |
| Zdroj: | Journal of Biological Chemistry. 268:19200-19203 |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/s0021-9258(19)36499-3 |
| Popis: | The rate-limiting step of eukaryotic protein synthesis is the binding of mRNA to the 40 S ribosomal subunit, a step which is catalyzed by initiation factors of the eIF-4 (eukaryotic initiation factor 4) group: eIF-4A, eIF-4B, eIF-4E, and eIF-4 gamma. Infection of cells with picornaviruses of the rhino- and enterovirus groups causes a shut-off in translation of cellular mRNAs but permits viral RNA translation to proceed. This change in translational specificity is thought to be mediated by proteolytic cleavage of eIF-4 gamma, which is catalyzed, directly or indirectly, by the picornaviral 2A protease. In this report we have used highly purified recombinant 2A protease from either human Coxsackievirus serotype B4 or rhinovirus serotype 2 to cleave eIF-4 gamma in vitro in the eIF-4 complex purified from rabbit reticulocytes. Neither the rate of cleavage nor fragment sizes were affected by addition of eIF-3. The NH2- and COOH-terminal fragments of eIF-4 gamma were separated by reverse phase HPLC and identified with specific antibodies, and the NH2-terminal sequence of the COOH-terminal fragment was determined by automated Edman degradation. The cleavage site for both proteases is 479GRPALSSR decreases GPPRGGPG494 in rabbit eIF-4 gamma, corresponding to 478GRTTLSTR decreases GPPRGGPG493 in human eIF-4 gamma. |
| Databáze: | OpenAIRE |
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