caa a22 4500 475766644 CHVBK 20180406123607.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1007/s007050070040 doi (NATIONALLICENCE)springer-10.1007/s007050070040 Mutations in a conserved enteroviral RNA oligonucleotide sequence affect positive strand viral RNA synthesis [Elektronische Daten] [S. Willian, S. Tracy, N. Chapman, S. Leser, J. Romero, B. Shapiro, K. Currey] Summary.:  We showed earlier that a transition mutation U234C, located within the completely conserved 5 nucleotide (nt) tract 5′-CGUUA (nt232-236) in the 5′ non-translated region (NTR) of the coxsackievirus B3 (CVB3) genome, attenuated CVB3 cardiovirulence in mice. To further explore the role of the sequence, we induced two single and one double transversion mutations in the conserved 5mer in a cardiovirulent CVB3 genome. The mutated sites partially or totally reverted to parental wild-type when progeny viruses were passaged at 37 °C, but remained stable when transfection and subsequent passages were performed at 33.5 °C. Viral replication in cell culture was attenuated at 37 °C or 39.5 °C relative to replication at 33.5 °C. While Western blot analysis demonstrated the level of protein translation consistent with virus replication, the ratios of positive to negative strand viral RNA at 37 °C in murine cells demonstrated a 2-5 fold diminution from those measured at 33.5 °C. Mutant CVB3 strains failed to replicate productively when inoculated into mice. The biological data are consistent with an hypothesis that proposes a lesion with primary effects at the level of positive strand viral RNA synthesis that results in attenuation of viral replication at physiologic temperature. Springer-Verlag/Wien, 2000 Willian S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut Tracy S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut Chapman N. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut Leser S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut Romero J. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut Shapiro B. Laboratory of Mathematical Biology, National Cancer Institute, FCRDC, DBS, Frederick, Maryland, U.S.A., US aut Currey K. Laboratory of Mathematical Biology, National Cancer Institute, FCRDC, DBS, Frederick, Maryland, U.S.A., US aut https://doi.org/10.1007/s007050070040 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s007050070040 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Willian S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut NATIONALLICENCE 700 1- Tracy S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut NATIONALLICENCE 700 1- Chapman N. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut NATIONALLICENCE 700 1- Leser S. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut NATIONALLICENCE 700 1- Romero J. Enterovirus Research Laboratory, Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska, U.S.A., US aut NATIONALLICENCE 700 1- Shapiro B. Laboratory of Mathematical Biology, National Cancer Institute, FCRDC, DBS, Frederick, Maryland, U.S.A., US aut NATIONALLICENCE 700 1- Currey K. Laboratory of Mathematical Biology, National Cancer Institute, FCRDC, DBS, Frederick, Maryland, U.S.A., US aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer