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Mutational analysis of the 3′‐terminal extra‐cistronic region of poliovirus RNA: secondary structure is not the only requirement for minus strand RNA replication
Author(s) -
A Pierangeli,
M Bucci,
P Pagnotti,
A M Degener,
R Perez Bercoff
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01127-z
Subject(s) - rna , biology , poliovirus , untranslated region , five prime untranslated region , rna editing , rna dependent rna polymerase , nucleic acid structure , non coding rna , microbiology and biotechnology , genetics , virology , gene , virus
A series of mutations were introduced in the 3′‐terminal untranslated region (3′‐UTR) of full‐length infectious poliovirus cDNA clones, and following transfection of COS‐1 cells the ability of these constructs to generate viable viral particles and/or to support viral RNA synthesis was assayed. Substitution of the 3′‐UTR of poliovirus RNA with the equivalent sequences of HAV RNA abrogated viral RNA replication, whereas the introduction of extended ‘foreign’ sequences between the open reading frame and the 3′‐UTR was well tolated. Point mutation that either destabilized the stem‐and‐loop structure or altered the sequence of the loop in domain ‘Y’ (nomenclature as per Pilipenko et al., [Nucleic Acids Res. 20 (1992) 1739–1745]) abolished both the infectivity and viral RNA synthesis. These were not restored by compensatory mutation that reconstructed the native secondary structure of this domain, suggesting that the secondary/tertiary folding of the 3′‐UTR is not the only determinant for template recognition at initiation of RNA synthesis, but rather that a specific primary sequence is indeed required.