+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment

Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment

Journal of General Virology 88(Pt 1): 342-350

The type member Mycoreovirus 1 (MyRV-1) of a newly described genus, Mycoreovirus, isolated from a hypovirulent strain 9B21 of the chestnut blight fungus, has a genome composed of 11 dsRNA segments (S1-S11). All of the segments have single ORFs on their capped, positive-sense strands. Infection of insect cells by baculovirus recombinants carrying full-length cDNAs of S1-S11 resulted in overexpression of protein products of the expected sizes, based on their deduced amino acid sequences. This expression system was utilized to identify the S3-encoded protein (VP3) as a guanylyltransferase by an autoguanylylation assay, in which only VP3 was radiolabelled with [alpha-(32)P]GTP. A series of progressive N-terminal and C-terminal deletion mutants was made to localize the autoguanylylation-active site of VP3 to aa residues 133-667. Within this region, a sequence stretch (aa 170-250) with relatively high sequence similarity to homologues of two other mycoreoviruses and two coltiviruses was identified. Site-directed mutagenesis of conserved aa residues revealed that H233, H242, Y243, F244 and F246, but not K172 or K202, play critical roles in guanylyltransferase activities. Together with broader sequence alignments of 'turreted' reoviruses, these results supported the a/vxxHx(8)Hyf/lvf motif, originally noted for orthoreovirus and aquareoviruses, as an active site for guanylyltransferases of viruses within the Orthoreovirus, Aquareovirus, Cypovirus, Oryzavirus, Fijivirus, Coltivirus and Mycoreovirus genera, as well as for the proposed Dinovernavirus genus.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 048353771

Download citation: RISBibTeXText

PMID: 17170467

Related references

Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment. Journal of General Virology 88(Part 1): 342-350, 2007

Unique establishment of procephalic head segments is supported by the identification of cis-regulatory elements driving segment-specific segment polarity gene expression in Drosophila. Development Genes and Evolution 221(1): 1-16, 2011

Genome segment 5 of Antheraea mylitta cytoplasmic polyhedrosis virus encodes a bona fide guanylyltransferase. Virology Journal 11: 53, 2014

Baculovirus expression of the small genome segment of Hantaan virus and potential use of the expressed nucleocapsid protein as a diagnostic antigen. Journal of General Virology 69: 777-786, 1988

Baculovirus expression of the M genome segment of Rift Valley fever virus and examination of antigenic and immunogenic properties of the expressed proteins. Virology 170(1): 184-192, 1989

Identification of genes encoding N-glycan processing beta-N-acetylglucosaminidases in Trichoplusia ni and Bombyx mori: Implications for glycoengineering of baculovirus expression systems. Biotechnology Progress 26(1): 34-44, 2010

Identification of multiple transcriptional initiation sites on the yeast mitochondrial genome by in vitro capping with guanylyltransferase. Journal of Biological Chemistry 258(22): 14025-14033, 1983

Guanylyltransferase activity of the LEF-4 subunit of baculovirus RNA polymerase. Journal of Virology 72(12): 10003-10010, 1998

Construction of chromosome segment substitution lines carrying overlapping chromosome segments of the whole wild rice genome and identification of quantitative trait loci for rice quality. Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao 32(3): 354-362, 2006

Nucleotide sequences of genome segments S8, encoding a capsid protein, and S10, encoding a 36K protein, of rice gall dwarf virus. Journal of General Virology 72: 2837-2842, 1991

Complete genome sequence of Mycoreovirus-1. Journal of general virology 85(11): 3437-3448, 2004

Capillary electrophoresis study on segment/segment system for segments based on phase of mixed micelles and its role in transport of particles between the two segments. Journal of Chromatography. a 1412: 139-150, 2015

Genome-wide identification, expression and chromosomal location of the genes encoding chitinolytic enzymes in Zea mays. Molecular Genetics and Genomics 280(2): 173-185, 2008

Mycoreovirus genome rearrangements associated with RNA silencing deficiency. Nucleic Acids Research 43(7): 3802-3813, 2015

Identification of signals required for the insertion of heterologous genome segments into the reovirus genome. Proceedings of the National Academy of Sciences of the United States of America 92(26): 12362-6, 1995