Genetic Evidence for an Interaction between a Picornaviral cis-Acting RNA Replication Element and 3CD Protein

Yan Yang, Rene Rijnbrand, Stanley Watowich, Stanley M. Lemon

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Internally located, cis-acting RNA replication elements, termed cres, are essential for replication of the genomes of picornaviruses such as human rhinovirus 14 (HRV-14) and poliovirus because they template uridylylation of the protein primer, VPg, by the polymerase 3Dpol. These cres form stem-loop structures sharing a common loop motif, and the HRV-14 cre can substitute functionally for the poliovirus cre in both uridylylation in vitro and RNA replication in vivo. We show, however, that the poliovirus cre is unable to support HRV-14 RNA replication. This lack of complementation maps to the stem of the poliovirus cre and was reversed by single nucleotide substitutions in the stem as well as the base of the loop. Replication-competent, revertant viruses rescued from dicistronic HRV-14 RNAs containing the poliovirus cre, or a chimeric cre containing the poliovirus stem, contained adaptive amino acid substitutions. These mapped to the surface of both the polymerase 3Dpol, at the tip of the "thumb" domain, and the protease 3Cpro, on the side opposing the active site and near the end of an extended strand segment implicated previously in RNA binding. These mutations substantially enhanced replication competence when introduced into HRV-14 RNAs containing the poliovirus cre, and they were additive in their effects. The data support a model in which 3CD or its derivatives 3Cpro and 3Dpol interact directly with the stem of the cre during uridylylation of VPg.

Original languageEnglish (US)
Pages (from-to)12659-12667
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number13
DOIs
StatePublished - Mar 26 2004

Fingerprint

Poliovirus
Rhinovirus
RNA
Proteins
Substitution reactions
Picornaviridae
Viruses
Thumb
Amino Acid Substitution
Mental Competency
Peptide Hydrolases
Nucleotides
Genes
Catalytic Domain
Derivatives
Amino Acids
Genome
Mutation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Genetic Evidence for an Interaction between a Picornaviral cis-Acting RNA Replication Element and 3CD Protein. / Yang, Yan; Rijnbrand, Rene; Watowich, Stanley; Lemon, Stanley M.

In: Journal of Biological Chemistry, Vol. 279, No. 13, 26.03.2004, p. 12659-12667.

Research output: Contribution to journalArticle

@article{7efe38fa22284914a6758b5d32172e18,
title = "Genetic Evidence for an Interaction between a Picornaviral cis-Acting RNA Replication Element and 3CD Protein",
abstract = "Internally located, cis-acting RNA replication elements, termed cres, are essential for replication of the genomes of picornaviruses such as human rhinovirus 14 (HRV-14) and poliovirus because they template uridylylation of the protein primer, VPg, by the polymerase 3Dpol. These cres form stem-loop structures sharing a common loop motif, and the HRV-14 cre can substitute functionally for the poliovirus cre in both uridylylation in vitro and RNA replication in vivo. We show, however, that the poliovirus cre is unable to support HRV-14 RNA replication. This lack of complementation maps to the stem of the poliovirus cre and was reversed by single nucleotide substitutions in the stem as well as the base of the loop. Replication-competent, revertant viruses rescued from dicistronic HRV-14 RNAs containing the poliovirus cre, or a chimeric cre containing the poliovirus stem, contained adaptive amino acid substitutions. These mapped to the surface of both the polymerase 3Dpol, at the tip of the {"}thumb{"} domain, and the protease 3Cpro, on the side opposing the active site and near the end of an extended strand segment implicated previously in RNA binding. These mutations substantially enhanced replication competence when introduced into HRV-14 RNAs containing the poliovirus cre, and they were additive in their effects. The data support a model in which 3CD or its derivatives 3Cpro and 3Dpol interact directly with the stem of the cre during uridylylation of VPg.",
author = "Yan Yang and Rene Rijnbrand and Stanley Watowich and Lemon, {Stanley M.}",
year = "2004",
month = "3",
day = "26",
doi = "10.1074/jbc.M312992200",
language = "English (US)",
volume = "279",
pages = "12659--12667",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "13",

}

TY - JOUR

T1 - Genetic Evidence for an Interaction between a Picornaviral cis-Acting RNA Replication Element and 3CD Protein

AU - Yang, Yan

AU - Rijnbrand, Rene

AU - Watowich, Stanley

AU - Lemon, Stanley M.

PY - 2004/3/26

Y1 - 2004/3/26

N2 - Internally located, cis-acting RNA replication elements, termed cres, are essential for replication of the genomes of picornaviruses such as human rhinovirus 14 (HRV-14) and poliovirus because they template uridylylation of the protein primer, VPg, by the polymerase 3Dpol. These cres form stem-loop structures sharing a common loop motif, and the HRV-14 cre can substitute functionally for the poliovirus cre in both uridylylation in vitro and RNA replication in vivo. We show, however, that the poliovirus cre is unable to support HRV-14 RNA replication. This lack of complementation maps to the stem of the poliovirus cre and was reversed by single nucleotide substitutions in the stem as well as the base of the loop. Replication-competent, revertant viruses rescued from dicistronic HRV-14 RNAs containing the poliovirus cre, or a chimeric cre containing the poliovirus stem, contained adaptive amino acid substitutions. These mapped to the surface of both the polymerase 3Dpol, at the tip of the "thumb" domain, and the protease 3Cpro, on the side opposing the active site and near the end of an extended strand segment implicated previously in RNA binding. These mutations substantially enhanced replication competence when introduced into HRV-14 RNAs containing the poliovirus cre, and they were additive in their effects. The data support a model in which 3CD or its derivatives 3Cpro and 3Dpol interact directly with the stem of the cre during uridylylation of VPg.

AB - Internally located, cis-acting RNA replication elements, termed cres, are essential for replication of the genomes of picornaviruses such as human rhinovirus 14 (HRV-14) and poliovirus because they template uridylylation of the protein primer, VPg, by the polymerase 3Dpol. These cres form stem-loop structures sharing a common loop motif, and the HRV-14 cre can substitute functionally for the poliovirus cre in both uridylylation in vitro and RNA replication in vivo. We show, however, that the poliovirus cre is unable to support HRV-14 RNA replication. This lack of complementation maps to the stem of the poliovirus cre and was reversed by single nucleotide substitutions in the stem as well as the base of the loop. Replication-competent, revertant viruses rescued from dicistronic HRV-14 RNAs containing the poliovirus cre, or a chimeric cre containing the poliovirus stem, contained adaptive amino acid substitutions. These mapped to the surface of both the polymerase 3Dpol, at the tip of the "thumb" domain, and the protease 3Cpro, on the side opposing the active site and near the end of an extended strand segment implicated previously in RNA binding. These mutations substantially enhanced replication competence when introduced into HRV-14 RNAs containing the poliovirus cre, and they were additive in their effects. The data support a model in which 3CD or its derivatives 3Cpro and 3Dpol interact directly with the stem of the cre during uridylylation of VPg.

UR - http://www.scopus.com/inward/record.url?scp=1842424947&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842424947&partnerID=8YFLogxK

U2 - 10.1074/jbc.M312992200

DO - 10.1074/jbc.M312992200

M3 - Article

C2 - 14711816

AN - SCOPUS:1842424947

VL - 279

SP - 12659

EP - 12667

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -