Specific interactions of HeLa cell proteins with proposed translation domains of the poliovirus 5' noncoding region.

Abstract:

:To determine which sequences or structures in the poliovirus 5' noncoding region (5'NCR) are involved in binding proteins used for internal ribosome binding and protein synthesis initiation, translation competition assays were performed in rabbit reticulocyte lysates in the presence and absence of HeLa cell extract. The results revealed two functional domains in the poliovirus 5'NCR. One, requiring nucleotides (nts) 457 to 626, binds proteins that are required for translation of all mRNAs and that are present in both reticulocyte lysates and HeLa cell extracts. Another, contained within nts 286 to 456, interacts with proteins that are specific for poliovirus translation and are present in HeLa cells but not in significant amounts in rabbit reticulocyte lysates. In order to detect HeLa cell proteins that interact stably with the 5'NCR of poliovirus, UV cross-linking was used. At least four major protein-RNA complexes were identified, three of which were shown by RNA competition analysis to bind specifically to defined domains within the 5'NCR. Protein A (54 kDa) cross-linked to RNA sequences and/or structures located between nts 457 and 626; proteins B (48 kDa) and C (38 kDa) bound to nts 286 to 456.

journal_name

J Virol

journal_title

Journal of virology

authors

Gebhard JR,Ehrenfeld E

doi

10.1128/JVI.66.5.3101-3109.1992

subject

Has Abstract

pub_date

1992-05-01 00:00:00

pages

3101-9

issue

5

eissn

0022-538X

issn

1098-5514

journal_volume

66

pub_type

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