DOI: 10.1128/jvi.76.9.4153-4161.2002. Both Viral Transcription and Replication Are Reduced when the Rabies Virus Nucleoprotein Is Not Phosphorylated

Both Viral Transcription and Replication Are Reduced when the Rabies Virus Nucleoprotein Is Not Phosphorylated

10.1128/jvi.76.9.4153-4161.2002

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

Abstract

ABSTRACT Rabies virus nucleoprotein (N) plays vital roles in regulation of viral RNA transcription and replication by encapsidation of the nascent genomic RNA. Rabies virus N is phosphorylated, and previous studies demonstrated that mutation of the phosphorylated serine at position 389 to alanine resulted in reduction of viral transcription and/or replication of a rabies virus minigenome. In the present study, we mutated the serine (S) at position 389 to alanine (A), glycine (G), aspartic acid (D), asparagine (N), glutamic acid (E), and glutamine (Q) and examined the effects of these mutations on rabies virus transcription and replication in the minigenome. Furthermore, mutations from S to A, S to D, and S to E were also incorporated into the full-length infectious virus. Mutation of the serine to each of the other amino acids resulted in the synthesis of an unphosphorylated N and reduction of viral transcription and replication in the minigenome. Mutations from S to A and S to D also resulted in reduction of both viral transcription and replication in full-length infectious viruses. Growth curve studies indicated that production of the mutant virus with the S-to-A mutation (L16A) was as much as 10,000-fold less than that of the wild-type virus (L16). Northern blot hybridization with rabies virus gene probes revealed that the rates of viral transcription and replication were reduced by as much as 10-fold in the mutant viruses when the N was not phosphorylated. Interpretation of the data from the minigenome system and the full-length infectious virus indicates that phosphorylation of rabies virus N is necessary for replication. Further studies involving cycloheximide treatment of infected cells revealed that viral transcription was also reduced when the N was not phosphorylated. Taken together, these results provide definitive evidence that N phosphorylation plays an important role in the processes of rabies virus transcription and replication.

Bibliography

Wu, X., Gong, X., Foley, H. D., Schnell, M. J., & Fu, Z. F. (2002). Both Viral Transcription and Replication Are Reduced when the Rabies Virus Nucleoprotein Is Not Phosphorylated. Journal of Virology, 76(9), 4153â4161.

Authors 5

Xianfu Wu (first)
Xiaoming Gong (additional)
Heather D. Foley (additional)
Matthias J. Schnell (additional)
Zhen F. Fu (additional)

References 28 Referenced 54

10.1016/S0065-3527(08)60860-X
10.1099/0022-1317-72-3-677
10.1006/fsim.2000.0326
10.1128/JVI.73.1.251-259.1999
Conzelmann, K.-K., J. Cox, L. G. Schneider, and H.-J. Theil. 1990. Molecular cloning and complete nucleotide sequence of the attenuated rabies virus SAD B19. J. Virol. 175 : 484-499. / J. Virol. (1990)
10.1128/jvi.68.2.713-719.1994
10.1016/0168-1702(87)90023-2
10.1016/0092-8674(82)90319-1
10.1128/jvi.71.10.7281-7288.1997
10.1073/pnas.88.5.2001
10.1016/0264-410X(93)90379-C
10.1089/oli.1.1996.6.87
10.1006/viro.1994.1222
10.1073/pnas.83.21.8122
10.1006/viro.1999.9962
10.1099/0022-1317-79-5-1005
10.1073/pnas.97.7.3544
10.1002/j.1460-2075.1994.tb06739.x
10.1056/NEJM199101243240401
10.1016/0042-6822(73)90413-3
10.1128/jvi.70.7.4538-4548.1996
10.1093/nar/14.6.2671
Wagner, R. R., and J. K. Rose. 1996. Rhabdoviridae: the viruses and their replication, p. 1121-1136. In B. N. Fields, D. M. Knipe, and P. M. Howley (ed.), Fields virology, 3rd ed. Lippincott-Raven Publishers, Philadelphia, Pa. / Fields virology (1996)
10.1016/0378-1119(94)90641-6
Wertz, G. W., N. L. Davies, and J. Patton. 1987. The role of proteins in vesicular stomatitis virus RNA replication, p. 271-296. In R. R. Wagner (ed.), The rhabdoviruses. Plenum Press, New York, N.Y. / The rhabdoviruses (1987)
10.1073/pnas.95.7.3501
Wunner, W. H. 1991. The chemical composition and molecular structure of rabies viruses, p. 31-67. In G. M. Baer (ed.), Natural history of rabies, 2nd ed. CRC Press, Inc., Boca Raton, Fla. / Natural history of rabies (1991)
10.1128/JVI.73.2.1661-1664.1999

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:59 a.m.)
Deposited	3 years, 5 months ago (March 5, 2022, 2:43 a.m.)
Indexed	1 month ago (Aug. 2, 2025, 12:06 a.m.)
Issued	23 years, 4 months ago (May 1, 2002)
Published	23 years, 4 months ago (May 1, 2002)
Published Print	23 years, 4 months ago (May 1, 2002)

Funders 0

None

BibTeX

@article{Wu_2002, title={Both Viral Transcription and Replication Are Reduced when the Rabies Virus Nucleoprotein Is Not Phosphorylated}, volume={76}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.76.9.4153-4161.2002}, DOI={10.1128/jvi.76.9.4153-4161.2002}, number={9}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Wu, Xianfu and Gong, Xiaoming and Foley, Heather D. and Schnell, Matthias J. and Fu, Zhen F.}, year={2002}, month=may, pages={4153–4161} }

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