Translation initiation in eukaryotes - chemical probing of encephalomyocarditis virus internal ribosomal entry siteстатья
Статья опубликована в журнале из списка RSCI Web of Science
Статья опубликована в журнале из перечня ВАК
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 25 апреля 2017 г.
Аннотация:A transcript including the internal ribosomal entry site (IRES, nts 315-833) and the beginning of the coding sequence of encephalomyocarditis virus (EMCV) RNA (nts 834-1155) was obtained using the TI polymerase system in vitro. A complex of this transcript with the 40S ribosomal subunit (48S preinitiation complex) was isolated from Krebs-2 mouse ascites carcinoma cell extracts. The complex was modified with dimethyl sulfate, which is widely used for chemical probing of the accessibility of A and C residues in single-stranded RNA regions. Modified nucleotides were identified by the primer extension inhibition analysis. The pattern of the 48S complex modification was compared with that of nonbound mRNP. It appeared that many A residues in IRES domains proximal to the initiation AUG codon (nts 834-836) were protected from modification. The data obtained are discussed in connection with the mechanism of internal translation initiation typical of EMCV and other picornaviral RNAs.