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Journal of Virology, January 1999, p. 805-809, Vol. 73, No. 1
Department of Microbiology and Immunology,
Wake Forest University School of Medicine, Winston-Salem, North
Carolina 27157-1064.
Received 7 July 1998/Accepted 16 September 1998
A functional RNA replication promoter for the paramyxovirus simian
virus 5 (SV5) requires two essential and discontinuous elements: 19 bases at the 3' terminus (conserved region I) and an 18-base internal
region (conserved region II [CRII]) that is contained within the
coding region of the L protein gene. A reverse-genetics system was used
to determine the sequence requirements for the internal CRII element to
function in RNA replication. A series of copyback defective interfering
(DI) RNA analogs were constructed to contain point mutations in
the 18 nucleotides composing CRII, and their relative replication
levels were analyzed. The results indicated that SV5 DI RNA replication
was reduced by substitutions for two CG dinucleotides, which in the
nucleocapsid template are in the first two positions of the first two
hexamers of CRII nucleotides. Substitutions for other bases within CRII
did not reduce RNA synthesis. Thus, two consecutive 5'-CGNNNN-3'
hexamers form an important sequence in the SV5 CRII promoter element.
The position of the CG dinucleotide within the SV5 leader and
antitrailer promoters was highly conserved among other members of the
Rubulavirus genus, but this motif differed significantly in
both sequence and position from that previously identified for Sendai
virus. The possible roles of the CRII internal promoter element in
paramyxovirus RNA replication are discussed.
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
RNA Replication for the Paramyxovirus Simian Virus 5 Requires
an Internal Repeated (CGNNNN) Sequence Motif
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Wake Forest University School of Medicine, Medical Center Blvd., Winston-Salem, NC 27157-1064. Phone: (336) 716-9083. Fax: (336) 716-9928. E-mail: gparks{at}wfubmc.edu.
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