The genomic RNA of Hepatitis A virus (HAV), a picornavirus of the hepatovirus group, is a single-stranded molecule, ca. 7.5 kb in length of positive polarity. Translation of this uncapped RNA starts at the 10th (or 11th) AUG triplet (position 734-36), by a mechanism of internal initiation of translation. The long sequences extending between the uncapped 5'-end and the translation initiation site contain two (instead of just one) pyrimidine-rich tracts (PRTs) spanning nucleotides 94-140 and 711-724, respectively. The latter lies only 11 nucleotides upstream from the initiation site of translation, and the question arose as to whether the notoriously poor replication ability of HAV was a consequence of a down regulation of translation due to the too short "spacer" sequence intervening between the 3'-PRT and the initiation of the main open reading frame. To address this issue, a series of full-length HAV cDNA clones were constructed in which the "spacer" sequence (normally 11 nts) was brought to 45 nts. Following transfection of COS-1 cells with these constructs, the amount of HAV (+)-strand RNA was determined by dot hybridization using a strand-specific RNA probe. HAV cDNA clones carrying a 45-nt "spacer" increased two-fold the rate of (+)-strand viral RNA synthesis, suggesting that the poor translation ability of HAV RNA may be one of the mechanisms responsible for the lengthy replication cycle of HAV.

INCREASED INFECTIVITY OF HAV CDNA CLONES WITH ENGINEERED 5'-TERMINAL EXTRA-CISTRONIC SEQUENCES / SILVEIRA CARNEIRO, J.; Bucci, Mauro; Pierangeli, Alessandra; Pagnotti, P.; Equestre, M.; PEREZ BERCOFF, Raul. - In: NEW MICROBIOLOGICA. - ISSN 1121-7138. - STAMPA. - 21:(1998), pp. 321-327.

INCREASED INFECTIVITY OF HAV CDNA CLONES WITH ENGINEERED 5'-TERMINAL EXTRA-CISTRONIC SEQUENCES.

BUCCI, Mauro;PIERANGELI, Alessandra;PEREZ BERCOFF, Raul
1998

Abstract

The genomic RNA of Hepatitis A virus (HAV), a picornavirus of the hepatovirus group, is a single-stranded molecule, ca. 7.5 kb in length of positive polarity. Translation of this uncapped RNA starts at the 10th (or 11th) AUG triplet (position 734-36), by a mechanism of internal initiation of translation. The long sequences extending between the uncapped 5'-end and the translation initiation site contain two (instead of just one) pyrimidine-rich tracts (PRTs) spanning nucleotides 94-140 and 711-724, respectively. The latter lies only 11 nucleotides upstream from the initiation site of translation, and the question arose as to whether the notoriously poor replication ability of HAV was a consequence of a down regulation of translation due to the too short "spacer" sequence intervening between the 3'-PRT and the initiation of the main open reading frame. To address this issue, a series of full-length HAV cDNA clones were constructed in which the "spacer" sequence (normally 11 nts) was brought to 45 nts. Following transfection of COS-1 cells with these constructs, the amount of HAV (+)-strand RNA was determined by dot hybridization using a strand-specific RNA probe. HAV cDNA clones carrying a 45-nt "spacer" increased two-fold the rate of (+)-strand viral RNA synthesis, suggesting that the poor translation ability of HAV RNA may be one of the mechanisms responsible for the lengthy replication cycle of HAV.
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INCREASED INFECTIVITY OF HAV CDNA CLONES WITH ENGINEERED 5'-TERMINAL EXTRA-CISTRONIC SEQUENCES / SILVEIRA CARNEIRO, J.; Bucci, Mauro; Pierangeli, Alessandra; Pagnotti, P.; Equestre, M.; PEREZ BERCOFF, Raul. - In: NEW MICROBIOLOGICA. - ISSN 1121-7138. - STAMPA. - 21:(1998), pp. 321-327.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/124843
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