Defective interfering (DI) RNAs of Berne virus (BEV) were generated by serial undiluted passaging of the virus in embryonic mule skin cells. Two DI RNAs of 1.0 and 1.4 kb (designated DI1000 and DI1400) were characterized in more detail. Isokinetic sucrose gradient analysis showed that these DI RNAs are specifically packaged into particles with smaller S values than standard virions. Both DI RNAs were cloned and sequenced. Three genomic cDNA clones were identified using probes complementary to the 5′ end of a DI RNA, which are thought to be derived from the 5′-terminal region of the BEV genome. A non-translated region of about 700 nt and the 5′ end of the putative BEV replicase gene were identified in the consensus nucleotide sequence. Both DI RNAs were shown to contain sequences from the 5′ and 3′ ends of the BEV genome. A conserved sequence motif, which has been postulated to be involved in sub-genomic RNA transcription, was also identified just downstream of the extreme 5′ ends of DI1000 and DI1400.
Den BoonJ. A.,
SnijderE. J.,
Krijnse LockerJ.,
HorzinekM. C.,
RottierP. J. M.1991; Another triple-spanning envelope protein among intracellularly budding RNA viruses: the torovirus E protein. Virology (in press)
HorzinekM. C.,
FlewettT. H.,
SaifL. J.,
SpaanW. J. M.,
WeissM.,
WoodeG. N.1987; A new family of vertebrate viruses: Toroviridae
. Intervirology 27:17–24
HuangA. S.1986; The role of defective interfering (DI) particles in viral infection. In The Molecular Basis of Viral Replication pp. 191–194 Edited by
Perez BercoffR.
New York: Plenum Press;
LevisR.,
WeissB. G.,
TsiangM.,
HuangH.,
SchlesingerS.1986; Deletion mapping of Sindbis virus DI RNAs derived from cDNAs defines the sequences essential for replication and packaging. Cell 44:137–145
MakinoS.,
ShiehC. K.,
BakerS. C.,
LaiM. M. C.1988; Primary structure and translation of defective interfering RNAs of murine coronavirus. Virology 166:550–560
PattnaikA. K.,
WertzG. W.1990; Replication and amplification of defective interfering particle RNAs of vesicular stomatitis virus in cells expressing viral proteins from vectors containing cloned cDNAs. Journal of Virology 64:2948–2957
SethnaP. B.,
HungS. L.,
BrianD. A.1989; Coronavirus subgenomic minus-strand RNAs and the potential for mRNA replicons. Proceedings of the National Academy of Sciences, U,. S,. A 86:5626–5630
SkinnerM. A.,
RacanielloV. R.,
DunnG.,
CooperJ.,
MinorP. D.,
AlmondJ. W.1989; New model for the secondary structure of the 5′ non-coding RNA of poliovirus is supported by biochemical and genetic data that also show that RNA secondary structure is important in neuroviolence. Journal of Molecular Biology 207:379–392
SnijderE. J.,
EderveenJ.,
SpaanW. J. M.,
WeissM.,
HorzinekM. C.1988; Characterization of Berne virus genomic and messenger RNAs. Journal of General Virology 69:2135–2144
SnijderE. J.,
Den BoonJ. A.,
BredenbeekP. J.,
HorzinekM. C.,
RijnbrandR.,
SpaanW. J. M.1990a; The carboxyl-terminal part of the putative Berne virus polymerase is expressed by ribosomal frameshifting and contains sequence motifs which indicate that toro- and coronaviruses are evolutionarily related. Nucleic Acids Research 18:4535–4542
SnijderE. J.,
Den BoonJ. A.,
SpaanW. J. M.,
WeissM.,
HorzinekM. C.1990b; Primary structure and post-translational processing of the Berne virus peplomer protein. Virology 178:35–363
SnijderE. J.,
HorzinekM. C.,
SpaanW. J. M.1990c; A 3′-coterminal nested set of independently transcribed messenger RNAs is generated during Berne virus replication. Journal of Virology 64:331–338
SnijderE. J.,
Den BoonJ. A.,
HorzinekM. C.,
SpaanW. J. M.1991; Comparison of the genome organization of toro- and coronaviruses: both divergence from a common ancestor and RNA recombination have played a role in Berne virus evolution. Virology 180:448–452
SonenbergN.1988; Cap-binding proteins of eukaryotic messenger RNAs: functions in initiation and control of translation. Progress in Nucleic Acids Research and Molecular Biology 35:173–207
Van der MostR. G.,
BredenbeerP. J.,
SpaanW. J. M.1991; A domain at the 3′ end of the polymerase gene is essential for encapsidation of coronavirus defective interfering RNAs. Journal of Virology 65: (in press)
Van der ZeijstB. A. M.,
BloemersH. P. J.1976; 4698 isokinetic glycerol and sucrose gradients for density gradient centrifugation. In Handbook of Biochemistry and Molecular Biology 3rd edn vol 1 pp. 426–519 Edited by
FasmanG. D.
Cleveland: CRC Press;
WeissM.,
SteckF.,
HorzinekM. C.1983; Purification and partial characterization of a new enveloped RNA virus (Berne virus). Journal of General Virology 64:1849–1858