Previous Article | Next Article 
Journal of Virology, February 1999, p. 1382-1391, Vol. 73, No. 2
0022-538X/99/$00.00+0
RNA-Binding and Capping Activities of Proteins
in Rotavirus Open Cores
John T.
Patton* and
Dayue
Chen
Laboratory of Infectious Diseases, National
Institute of Allergy and Infectious Diseases, National Institutes
of Health, Bethesda, Maryland 20892
Received 27 May 1998/Accepted 5 November 1998
Guanylyltransferases are members of the nucleotidyltransferase
family and function in mRNA capping by transferring GMP to the
phosphate end of nascent RNAs. Although numerous guanylyltransferases have been identified, studies which define the nature of the
interaction between the capping enzymes of any origin and their RNA
substrates have been limited. Here, we have characterized the
RNA-binding activity of VP3, a minor protein component of the core of
rotavirions that has been proposed to function as the viral
guanylyltransferase and to direct the capping of the 11 transcripts
synthesized from the segmented double-stranded RNA (dsRNA) genome of
these viruses. Gel shift analysis performed with disrupted (open)
virion-derived cores and virus-specific RNA probes showed that VP3 has
affinity for single-stranded RNA (ssRNA) but not for dsRNA. While the
ssRNA-binding activity of VP3 was found to be sequence independent, the
protein does exhibit preferential affinity for uncapped over capped
RNA. Like the RNA-binding activity, RNA capping assays performed with open cores indicates that the guanylyltransferase activity of VP3 is
nonspecific and is able to cap RNAs initiating with a G or an A
residue. These data establish that all three rotavirus core proteins,
VP1, the RNA polymerase; VP2, the core capsid protein; and VP3, the
guanylyltransferase, have affinity for RNA but that only in the case of
the RNA polymerase is the affinity sequence specific.
*
Corresponding author. Mailing address: Laboratory of
Infectious Diseases, National Institute of Allergy and Infectious
Diseases, National Institutes of Health, 7 Center Dr., MSC 0720, Rm.
117, Bethesda, MD 20892. Phone: (301) 496-3372. Fax: (301) 496-8312. E-mail: JPATTON{at}atlas.niaid.nih.gov.
Journal of Virology, February 1999, p. 1382-1391, Vol. 73, No. 2
0022-538X/99/$00.00+0
This article has been cited by other articles:
-
Ayala-Breton, C., Arias, M., Espinosa, R., Romero, P., Arias, C. F., Lopez, S.
(2009). Analysis of the Kinetics of Transcription and Replication of the Rotavirus Genome by RNA Interference. J. Virol.
83: 8819-8831
[Abstract]
[Full Text]
-
Roy, P.
(2008). Bluetongue virus: dissection of the polymerase complex. J. Gen. Virol.
89: 1789-1804
[Abstract]
[Full Text]
-
TORTORICI, M. A., SHAPIRO, B. A., PATTON, J. T.
(2006). A base-specific recognition signal in the 5' consensus sequence of rotavirus plus-strand RNAs promotes replication of the double-stranded RNA genome segments. RNA
12: 133-146
[Abstract]
[Full Text]
-
Campagna, M., Eichwald, C., Vascotto, F., Burrone, O. R.
(2005). RNA interference of rotavirus segment 11 mRNA reveals the essential role of NSP5 in the virus replicative cycle. J. Gen. Virol.
86: 1481-1487
[Abstract]
[Full Text]
-
Jaafar, F. M., Attoui, H., Mertens, P. P. C., de Micco, P., de Lamballerie, X.
(2005). Identification and functional analysis of VP3, the guanylyltransferase of Banna virus (genus Seadornavirus, family Reoviridae). J. Gen. Virol.
86: 1141-1146
[Abstract]
[Full Text]
-
Boyce, M., Wehrfritz, J., Noad, R., Roy, P.
(2004). Purified Recombinant Bluetongue Virus VP1 Exhibits RNA Replicase Activity. J. Virol.
78: 3994-4002
[Abstract]
[Full Text]
-
Tortorici, M. A., Broering, T. J., Nibert, M. L., Patton, J. T.
(2003). Template Recognition and Formation of Initiation Complexes by the Replicase of a Segmented Double-stranded RNA Virus. J. Biol. Chem.
278: 32673-32682
[Abstract]
[Full Text]
-
Taraporewala, Z. F., Schuck, P., Ramig, R. F., Silvestri, L., Patton, J. T.
(2002). Analysis of a Temperature-Sensitive Mutant Rotavirus Indicates that NSP2 Octamers Are the Functional Form of the Protein. J. Virol.
76: 7082-7093
[Abstract]
[Full Text]
-
Vende, P., Taraporewala, Z. F., Patton, J. T.
(2002). RNA-Binding Activity of the Rotavirus Phosphoprotein NSP5 Includes Affinity for Double-Stranded RNA. J. Virol.
76: 5291-5299
[Abstract]
[Full Text]
-
Taraporewala, Z. F., Patton, J. T.
(2001). Identification and Characterization of the Helix-Destabilizing Activity of Rotavirus Nonstructural Protein NSP2. J. Virol.
75: 4519-4527
[Abstract]
[Full Text]
-
Lawton, J. A., Estes, M. K., Prasad, B. V. V.
(2001). Identification and Characterization of a Transcription Pause Site in Rotavirus. J. Virol.
75: 1632-1642
[Abstract]
[Full Text]
-
Taraporewala, Z., Chen, D., Patton, J. T.
(1999). Multimers Formed by the Rotavirus Nonstructural Protein NSP2 Bind to RNA and Have Nucleoside Triphosphatase Activity. J. Virol.
73: 9934-9943
[Abstract]
[Full Text]