Rescue of a bovine respiratory syncytial virus genomic RNA analog by bovine, human and ovine respiratory syncytial viruses confirms the “functional integrity” and “cross-recognition” of BRSV cis-acting elements by HRSV and ORSV

The nucleotide sequences of the 3′ leader and 5′ trailer regions were determined for genomic RNA of bovine respiratory syncytial virus (BRSV) strain A-51908. The leader and trailer sequences are ‘45’ and ‘161’ nucleotides in length, respectively. The functionality of BRSV leader and trailer sequence...

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Veröffentlicht in:Archives of virology 1999-01, Vol.144 (10), p.1977-1990
Hauptverfasser: Yunus, A. S, Krishnamurthy, S, Pastey, M. K, Huang, Z, Khattar, S. K, Collins, P. L, Samal, S. K
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container_end_page 1990
container_issue 10
container_start_page 1977
container_title Archives of virology
container_volume 144
creator Yunus, A. S
Krishnamurthy, S
Pastey, M. K
Huang, Z
Khattar, S. K
Collins, P. L
Samal, S. K
description The nucleotide sequences of the 3′ leader and 5′ trailer regions were determined for genomic RNA of bovine respiratory syncytial virus (BRSV) strain A-51908. The leader and trailer sequences are ‘45’ and ‘161’ nucleotides in length, respectively. The functionality of BRSV leader and trailer sequences and their recognition by HRSV and ovine respiratory syncytial virus (ORSV) proteins were examined with a in vitro transcribed BRSV genomic RNA analog carrying the bacterial chloramphenicol acetyl transferase (CAT) gene under the control of BRSV transcription signals. Upon transfection into BRSV, HRSV or ORSV infected cells, the BRSV minireplicons were ‘rescued’ such that the reporter gene was expressed, the minigenome was replicated and packaged into micrococcal nuclease resistant-infectious minireplicons. The passage of infectious minireplicons could be blocked by a polyclonal BRSV neutralizing antiserum. Bovine parainfluenza virus-3, a heterologous paramyxovirus was inactive in rescuing BRSV genomic RNA analog. Mutational substitution of the G residue at position 4 of leader sequence in the BRSV genomic RNA analog, with an A or U residue inhibited its transcription and replication, while replacement with a C residue had no significant effect on rescue. These results show that the cis-acting elements of BRSV are functional and are also recognized by the proteins of HRSV and ORSV. The helper virus complemented rescue system developed here will be useful for characterizing the cis-acting elements of BRSV.
doi_str_mv 10.1007/s007050050719
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subjects 3' Untranslated Regions - genetics
5' Untranslated Regions - genetics
Animals
antiserum
Biological and medical sciences
Blotting, Northern
Bovine respiratory syncytial virus
Cattle
chloramphenicol acetyltransferase
Chloramphenicol O-Acetyltransferase - genetics
Chloramphenicol O-Acetyltransferase - metabolism
Cloning, Molecular
DNA, Complementary - genetics
Fundamental and applied biological sciences. Psychology
Genetics
Genome, Viral
Helper Viruses - genetics
Helper Viruses - metabolism
Human respiratory syncytial virus
Humans
Microbiology
micrococcal nuclease
Molecular Sequence Data
Mutation
neutralization
Neutralization Tests
nucleotide sequences
Ovine respiratory syncytial virus
proteins
Replicon
reporter genes
Respiratory syncytial virus
Respiratory Syncytial Virus, Bovine - genetics
Respiratory Syncytial Virus, Bovine - physiology
Respiratory Syncytial Virus, Human - genetics
Respiratory Syncytial Viruses - genetics
Respirovirus
RNA
RNA, Viral - genetics
RNA, Viral - isolation & purification
sheep
Transcription, Genetic
Transfection
Virology
Virus Replication
viruses
title Rescue of a bovine respiratory syncytial virus genomic RNA analog by bovine, human and ovine respiratory syncytial viruses confirms the “functional integrity” and “cross-recognition” of BRSV cis-acting elements by HRSV and ORSV
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