Structure and Expression of the Picornavirus Genome*

Kitamura, Naomi; Adler, Cheryl J.; Wimmer, Eckard

doi:10.1111/j.1749-6632.1980.tb27967.x

Cited by 40 publications

(26 citation statements)

References 77 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In a possibly analogous fashion, adenovirus DNA synthesis may be facilitated by the covalent complex formed between the virus 5'-terminal protein and dCMP, the nucleotide that initiates DNA replication (9). The same type of nucleotidyl-protein complex may function in genome replication of other DNA viruses (6,13) and some RNA viruses (7) and perhaps in uninfected cells as well (2).…”

Section: Methodsmentioning

confidence: 99%

Covalent guanylyl intermediate formed by HeLa cell mRNA capping enzyme.

Wang¹,

Furuichi²,

Shatkin³

1982

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Covalent guanylyl intermediate formed by HeLa cell mRNA capping enzyme.

Wang¹,

Furuichi²,

Shatkin³

1982

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

“…Virus-specific mRNA, which differs from genome RNA only at its 5' terminus (1)(2)(3), is translated to produce one large polyprotein that is subsequently processed by proteolytic cleavage into smaller, functional proteins (4-7). The nucleotide sequence of the poliovirus genome has a long reading frame encompassing 89% of the RNA (8,9), an observation in accordance with polyprotein synthesis.…”

mentioning

confidence: 99%

Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3-7c inhibit cleavage at glutamine-glycine pairs.

Hanecak¹,

Semler²,

Anderson³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

205

143

View full text Add to dashboard Cite

Proteolytic processing of poliovirus polypeptides was examined by the addition of antibodies directed against the viral proteins P3-7c and P2-X to a cell-free translation extract prepared from infected HeLa cells. Antisera to P3-7c specifically inhibited in vitro processing at Gln-Gly pairs. Partial amino acid sequence analysis revealed a second Tyr-Gly pair that is utilized in protein processing. Neither Tyr-Gly cleavage is affected by antibody to P3-7c. Anti-P3-7c antibodies react not only with P3-7c but also with P3-6a and P3-2, two viral polypeptides NH2-coterminal with P3-7c. Preimmune and anti-P2-X antibodies had no effect on the processing of poliovirus proteins in vitro. We conclude that the activity responsible for processing poliovirus polypeptides at Gln-Gly pairs resides in the primary structure of P3-7c (or P3-2 and P3-6a) and not in P2-X.

show abstract

“…Its 5' end is covalently attached to a small viral protein (VP) termed VPg (for review, see ref. 1). The complete sequence of poliovirus type 1 (PV-1) RNA has recently been determined (2).…”

mentioning

confidence: 99%

Molecular cloning of the genome of poliovirus type 1.

Werf¹,

Brégégère²,

Kopecká³

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Poliovirus cDNA RNA hybrids were prepared from the Mahoney strain ofpoliovirus type 1 by using reverse transcriptase (RNA-dependent DNA nucleotidyltransferase) and cloned in the Escherichia coliplasmidpBR322. Bacteria colonies carrying recombinant plasmids were selected by in situ hybridization with virus-specific RNase Ti-resistant oligonucleotides. Analysis of the cDNA inserts by restriction mapping and electron microscopy showed that the cloned cDNAs, the longest of which was 3.2 kIlobase pairs, originated from various parts of the viral RNA, covering at least 99% ofthe genome length. Due to overlapping ofthe clones, the restriction map of the poliovirus genome could be reconstructed. The complete 5' end of the genome was successfully cloned in at least one of the recombinant plasmids, pPVl-366.The genome of poliovirus is a polyadenylylated RNA of plusstrand polarity. Its 5' end is covalently attached to a small viral protein (VP) termed VPg (for review, see ref. 1). The complete sequence of poliovirus type 1 (PV-1) RNA has recently been determined (2). Poliovirus has a unique strategy ofgene expression; its primary translation product is a 220,000-dalton polyprotein, noncapsid VP (NCVP) NCVP00, that is processed into numerous polypeptides by proteinase(s) (3). Most of the viral polypeptides have been mapped; the capsid proteins are encoded in the 5'-terminal halfofthe genome in the order 5'-VP4-VP2-VP3-VP1, whereas nonstructural proteins such as NCVP X, a putative proteinase, and NCVP4, the viral RNA polymerase (4, 5), are encoded in the middle and the 3' part of the genome, respectively (2, 3). However, the functions of many viral polypeptides are still obscure. Also, little is known about the regulation of proteolytic processing or the mechanism of RNA replication.Recombinant DNA technology allows synthesis and cloning in Escherichia coli of double-stranded cDNA copies of a viral RNA genome. The availability of such clones would be helpful for detailed analysis of the organization of the poliovirus genome, including confirmation of the nucleotide sequence information obtained by other means. In addition, the expression of virus-specific proteins in E. coli may open the way to the development of new types of vaccine. This is desirable in view of the fact that vaccination with both killed and live poliovirus has met with difficulties, particularly in tropical countries. Finally, should expression of cloned poliovirus cDNA in eukaryotic cells prove possible, the system would offer a unique opportunity for in vitro site-directed mutagenesis and thus for genetic analyses of the virus.In this paper, we report the isolation of PV-1 cDNA clones that together cover 99% of the viral genome, and we present a restriction map of poliovirus-specific DNA. MATERIALS AND METHODSConstruction and Screening of Clones Containing Recombinant DNAs. The Mahoney strain of PV-1 was grown in suspension cultures of HeLa cells, and viral RNA was extracted as described (6). cDNA was prepared by using an oligo(dT)10 primer and ...

show abstract

Structure and Expression of the Picornavirus Genome*

Cited by 40 publications

References 77 publications

Covalent guanylyl intermediate formed by HeLa cell mRNA capping enzyme.

Covalent guanylyl intermediate formed by HeLa cell mRNA capping enzyme.

Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3-7c inhibit cleavage at glutamine-glycine pairs.

Molecular cloning of the genome of poliovirus type 1.

Contact Info

Product

Resources

About