Analysis of efficiently packaged defective interfering RNAs of murine coronavirus

Localization of a possible RNA-packaging signal

Shinji Makino, Kyoko Yokomori, Michael M C Lai

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

We have previously shown that most of the defective interfering (DI) RNA of mouse hepatitis (MHV) are not packaged into virions. We have now identified, after 21 serial undiluted of MHV, a small DI RNA, DIssF, which is efficiently packaged into virions. The DIssF RNA replicated at a on its transfection into the helper virus-infected cells. The virus released from the transfected cells interfered strongly with mRNA synthesis and growth of helper virus. cDNA of cloning and sequence analysis of DIssF RNA revealed the that it is 3.6 kb and consists of sequences derived from five discontinuous regions of the genome of the nondefective virus. The first four regions (domains I to IV) from the 5′ end are derived from gene 1, which presumably encodes the RNA polymerase of the nondefective virus. The entire domain I (859 nucleotides) and the first 750 nucleotides of domain II are also present in a previously characterized DI RNA, DIssE, which is not efficiently packaged into virions. Furthermore, the junction between these two domains is identical between the two DI RNAs. The remaining 77 nucleotides at the 3′ of II all of domains III (655 nucleotides) and IV (770 nucleotides) are not present in DIssE RNA. These four domains are derived from gene 1. In contrast the 3′-most domain (domain V, 447 nucleotides) is derived from the 3′ end of the genomic RNA and is also present in DIssE. The comparison of primary sequences and packaging properties between DIssE and DIssF RNAs suggested that domains III and IV and part of the 3′ end of domain II contain the packaging signal for MHV RNA. This conclusion was confirmed by inserting these DIssF-unique sequences into a DIssE cDNA construct; the in vitro-transcribed RNA from this hybrid construct was efficiently into virion particles. DIssF RNA also contains an open reading frame, which begins from domain I and ends at the 5′-end 20 bases of domain III. In vitro translation of DIssF RNA and metabolic labeling of the virus-infected cells showed that this open reading frame is indeed translated into a 74-kDa protein. The structures of both DIssE and DIssF RNAs suggest that a protein-encoding capability is a common characteristic of MHV DI RNA.

Original languageEnglish (US)
Pages (from-to)6045-6053
Number of pages9
JournalJournal of Virology
Volume64
Issue number12
StatePublished - 1990
Externally publishedYes

Fingerprint

Murine coronavirus
Coronavirus
Product Packaging
RNA
Nucleotides
nucleotides
Virion
virion
viruses
Helper Viruses
Viruses
RNA packaging
Open Reading Frames
packaging
open reading frames
Complementary DNA
RNA Sequence Analysis

ASJC Scopus subject areas

  • Immunology

Cite this

Analysis of efficiently packaged defective interfering RNAs of murine coronavirus : Localization of a possible RNA-packaging signal. / Makino, Shinji; Yokomori, Kyoko; Lai, Michael M C.

In: Journal of Virology, Vol. 64, No. 12, 1990, p. 6045-6053.

Research output: Contribution to journalArticle

@article{898beb6c69fe402bb7dd70bc21dfad13,
title = "Analysis of efficiently packaged defective interfering RNAs of murine coronavirus: Localization of a possible RNA-packaging signal",
abstract = "We have previously shown that most of the defective interfering (DI) RNA of mouse hepatitis (MHV) are not packaged into virions. We have now identified, after 21 serial undiluted of MHV, a small DI RNA, DIssF, which is efficiently packaged into virions. The DIssF RNA replicated at a on its transfection into the helper virus-infected cells. The virus released from the transfected cells interfered strongly with mRNA synthesis and growth of helper virus. cDNA of cloning and sequence analysis of DIssF RNA revealed the that it is 3.6 kb and consists of sequences derived from five discontinuous regions of the genome of the nondefective virus. The first four regions (domains I to IV) from the 5′ end are derived from gene 1, which presumably encodes the RNA polymerase of the nondefective virus. The entire domain I (859 nucleotides) and the first 750 nucleotides of domain II are also present in a previously characterized DI RNA, DIssE, which is not efficiently packaged into virions. Furthermore, the junction between these two domains is identical between the two DI RNAs. The remaining 77 nucleotides at the 3′ of II all of domains III (655 nucleotides) and IV (770 nucleotides) are not present in DIssE RNA. These four domains are derived from gene 1. In contrast the 3′-most domain (domain V, 447 nucleotides) is derived from the 3′ end of the genomic RNA and is also present in DIssE. The comparison of primary sequences and packaging properties between DIssE and DIssF RNAs suggested that domains III and IV and part of the 3′ end of domain II contain the packaging signal for MHV RNA. This conclusion was confirmed by inserting these DIssF-unique sequences into a DIssE cDNA construct; the in vitro-transcribed RNA from this hybrid construct was efficiently into virion particles. DIssF RNA also contains an open reading frame, which begins from domain I and ends at the 5′-end 20 bases of domain III. In vitro translation of DIssF RNA and metabolic labeling of the virus-infected cells showed that this open reading frame is indeed translated into a 74-kDa protein. The structures of both DIssE and DIssF RNAs suggest that a protein-encoding capability is a common characteristic of MHV DI RNA.",
author = "Shinji Makino and Kyoko Yokomori and Lai, {Michael M C}",
year = "1990",
language = "English (US)",
volume = "64",
pages = "6045--6053",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Analysis of efficiently packaged defective interfering RNAs of murine coronavirus

T2 - Localization of a possible RNA-packaging signal

AU - Makino, Shinji

AU - Yokomori, Kyoko

AU - Lai, Michael M C

PY - 1990

Y1 - 1990

N2 - We have previously shown that most of the defective interfering (DI) RNA of mouse hepatitis (MHV) are not packaged into virions. We have now identified, after 21 serial undiluted of MHV, a small DI RNA, DIssF, which is efficiently packaged into virions. The DIssF RNA replicated at a on its transfection into the helper virus-infected cells. The virus released from the transfected cells interfered strongly with mRNA synthesis and growth of helper virus. cDNA of cloning and sequence analysis of DIssF RNA revealed the that it is 3.6 kb and consists of sequences derived from five discontinuous regions of the genome of the nondefective virus. The first four regions (domains I to IV) from the 5′ end are derived from gene 1, which presumably encodes the RNA polymerase of the nondefective virus. The entire domain I (859 nucleotides) and the first 750 nucleotides of domain II are also present in a previously characterized DI RNA, DIssE, which is not efficiently packaged into virions. Furthermore, the junction between these two domains is identical between the two DI RNAs. The remaining 77 nucleotides at the 3′ of II all of domains III (655 nucleotides) and IV (770 nucleotides) are not present in DIssE RNA. These four domains are derived from gene 1. In contrast the 3′-most domain (domain V, 447 nucleotides) is derived from the 3′ end of the genomic RNA and is also present in DIssE. The comparison of primary sequences and packaging properties between DIssE and DIssF RNAs suggested that domains III and IV and part of the 3′ end of domain II contain the packaging signal for MHV RNA. This conclusion was confirmed by inserting these DIssF-unique sequences into a DIssE cDNA construct; the in vitro-transcribed RNA from this hybrid construct was efficiently into virion particles. DIssF RNA also contains an open reading frame, which begins from domain I and ends at the 5′-end 20 bases of domain III. In vitro translation of DIssF RNA and metabolic labeling of the virus-infected cells showed that this open reading frame is indeed translated into a 74-kDa protein. The structures of both DIssE and DIssF RNAs suggest that a protein-encoding capability is a common characteristic of MHV DI RNA.

AB - We have previously shown that most of the defective interfering (DI) RNA of mouse hepatitis (MHV) are not packaged into virions. We have now identified, after 21 serial undiluted of MHV, a small DI RNA, DIssF, which is efficiently packaged into virions. The DIssF RNA replicated at a on its transfection into the helper virus-infected cells. The virus released from the transfected cells interfered strongly with mRNA synthesis and growth of helper virus. cDNA of cloning and sequence analysis of DIssF RNA revealed the that it is 3.6 kb and consists of sequences derived from five discontinuous regions of the genome of the nondefective virus. The first four regions (domains I to IV) from the 5′ end are derived from gene 1, which presumably encodes the RNA polymerase of the nondefective virus. The entire domain I (859 nucleotides) and the first 750 nucleotides of domain II are also present in a previously characterized DI RNA, DIssE, which is not efficiently packaged into virions. Furthermore, the junction between these two domains is identical between the two DI RNAs. The remaining 77 nucleotides at the 3′ of II all of domains III (655 nucleotides) and IV (770 nucleotides) are not present in DIssE RNA. These four domains are derived from gene 1. In contrast the 3′-most domain (domain V, 447 nucleotides) is derived from the 3′ end of the genomic RNA and is also present in DIssE. The comparison of primary sequences and packaging properties between DIssE and DIssF RNAs suggested that domains III and IV and part of the 3′ end of domain II contain the packaging signal for MHV RNA. This conclusion was confirmed by inserting these DIssF-unique sequences into a DIssE cDNA construct; the in vitro-transcribed RNA from this hybrid construct was efficiently into virion particles. DIssF RNA also contains an open reading frame, which begins from domain I and ends at the 5′-end 20 bases of domain III. In vitro translation of DIssF RNA and metabolic labeling of the virus-infected cells showed that this open reading frame is indeed translated into a 74-kDa protein. The structures of both DIssE and DIssF RNAs suggest that a protein-encoding capability is a common characteristic of MHV DI RNA.

UR - http://www.scopus.com/inward/record.url?scp=0025205237&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025205237&partnerID=8YFLogxK

M3 - Article

VL - 64

SP - 6045

EP - 6053

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 12

ER -