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Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA.
RNA. 1995 Nov; 1(9):924-38.RNA

Abstract

The requirement of PTB, polypyrimidine tract binding protein, for internal initiation of translation has been tested using an RNA affinity column to deplete rabbit reticulocyte lysates of PTB. The affinity column was prepared by coupling CNBr-activated Sepharose with the segment of the 5'-untranslated region of encephalomyocarditis virus (EMCV) RNA previously shown to bind PTB. Lysates passed through this column were devoid of PTB, and were incapable of internal initiation of translation dependent on the EMCV 5'-untranslated region, while retaining the capacity for translation dependent on ribosome scanning. Full activity for internal initiation was restored by the addition of recombinant PTB at the physiologically relevant concentration of about 5 micrograms/mL. When various PTB deletion mutants were tested, it was found that this activity required virtually the full-length protein. Thus, PTB is an essential protein for internal initiation promoted by the EMCV 5'-untranslated region. However, the PTB-depleted lysate retained the capacity for internal initiation promoted by the 5'-untranslated regions of another cardiovirus, Theiler's murine encephalomyelitis virus, and of the unrelated hepatitis C virus, and in neither case did addition of recombinant PTB stimulate internal initiation. Therefore, PTB is not a universal internal initiation factor that is indispensable in every case of internal ribosome entry.

Authors+Show Affiliations

Department of Biochemistry, University of Cambridge, United Kingdom.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8548657

Citation

Kaminski, A, et al. "Direct Evidence That Polypyrimidine Tract Binding Protein (PTB) Is Essential for Internal Initiation of Translation of Encephalomyocarditis Virus RNA." RNA (New York, N.Y.), vol. 1, no. 9, 1995, pp. 924-38.
Kaminski A, Hunt SL, Patton JG, et al. Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA. RNA. 1995;1(9):924-38.
Kaminski, A., Hunt, S. L., Patton, J. G., & Jackson, R. J. (1995). Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA. RNA (New York, N.Y.), 1(9), 924-38.
Kaminski A, et al. Direct Evidence That Polypyrimidine Tract Binding Protein (PTB) Is Essential for Internal Initiation of Translation of Encephalomyocarditis Virus RNA. RNA. 1995;1(9):924-38. PubMed PMID: 8548657.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Direct evidence that polypyrimidine tract binding protein (PTB) is essential for internal initiation of translation of encephalomyocarditis virus RNA. AU - Kaminski,A, AU - Hunt,S L, AU - Patton,J G, AU - Jackson,R J, PY - 1995/11/1/pubmed PY - 1995/11/1/medline PY - 1995/11/1/entrez SP - 924 EP - 38 JF - RNA (New York, N.Y.) JO - RNA VL - 1 IS - 9 N2 - The requirement of PTB, polypyrimidine tract binding protein, for internal initiation of translation has been tested using an RNA affinity column to deplete rabbit reticulocyte lysates of PTB. The affinity column was prepared by coupling CNBr-activated Sepharose with the segment of the 5'-untranslated region of encephalomyocarditis virus (EMCV) RNA previously shown to bind PTB. Lysates passed through this column were devoid of PTB, and were incapable of internal initiation of translation dependent on the EMCV 5'-untranslated region, while retaining the capacity for translation dependent on ribosome scanning. Full activity for internal initiation was restored by the addition of recombinant PTB at the physiologically relevant concentration of about 5 micrograms/mL. When various PTB deletion mutants were tested, it was found that this activity required virtually the full-length protein. Thus, PTB is an essential protein for internal initiation promoted by the EMCV 5'-untranslated region. However, the PTB-depleted lysate retained the capacity for internal initiation promoted by the 5'-untranslated regions of another cardiovirus, Theiler's murine encephalomyelitis virus, and of the unrelated hepatitis C virus, and in neither case did addition of recombinant PTB stimulate internal initiation. Therefore, PTB is not a universal internal initiation factor that is indispensable in every case of internal ribosome entry. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/8548657/Direct_evidence_that_polypyrimidine_tract_binding_protein__PTB__is_essential_for_internal_initiation_of_translation_of_encephalomyocarditis_virus_RNA_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=8548657 DB - PRIME DP - Unbound Medicine ER -