Direct characterization of influenza viral NS1 mRNA and related sequences from infected HeLa cells and a cell-free transcription system

Allen W. Nicholson; Howard M. Frankfort; Nicholas G. Davis; Stefano Ferrari; Robert A. Lamb; Hugh D. Robertson

doi:10.1016/0167-4781(86)90018-7

Direct characterization of influenza viral NS₁ mRNA and related sequences from infected HeLa cells and a cell-free transcription system

Allen W. Nicholson^*, Howard M. Frankfort, Nicholas G. Davis, Stefano Ferrari, Robert A. Lamb, Hugh D. Robertson

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

Abstract

The NS₁ mRNA of the influenza A virus WSN (H₀N₁) strain was isolated from a cell-free transcription system, and from the cytoplasm of virus-infected HeLa cells. The ³²P-labeled NS₁ mRNA derived from the infected cell cytoplasm was characterized by the secondary enzymatic analysis of sixteen of its large or distinct RNAase T1-resistant oligonucleotides. Several WSN strain-specific nucleotide differences from the previously-determined sequence of NS₁ mRNA from the PR8 (H₀N₁) strain of influenza A virus, were located within these sequences. The RNAase T1-resistant oligonucleotides were placed within the primary sequence of NS₁ mRNA, using the PR8 strain sequence data. The resulting linear map was then used to identify NS₂ mRNA isolated from the infected cell cytoplasm, and an NS-related RNA species generated from NS₁ mRNA incubated in a HeLa cell-free extract.

Original language	English (US)
Pages (from-to)	153-163
Number of pages	11
Journal	BBA - Gene Structure and Expression
Volume	868
Issue number	2-3
DOIs	https://doi.org/10.1016/0167-4781(86)90018-7
State	Published - Nov 13 1986

Keywords

Influenza mRNA
Primed RNA synthesis
RNA processing
mRNA

ASJC Scopus subject areas

Structural Biology
Biophysics
Biochemistry
Genetics

Access to Document

10.1016/0167-4781(86)90018-7

Cite this

@article{704ff289f496462eab46f2b765d54ff3,

title = "Direct characterization of influenza viral NS1 mRNA and related sequences from infected HeLa cells and a cell-free transcription system",

abstract = "The NS1 mRNA of the influenza A virus WSN (H0N1) strain was isolated from a cell-free transcription system, and from the cytoplasm of virus-infected HeLa cells. The 32P-labeled NS1 mRNA derived from the infected cell cytoplasm was characterized by the secondary enzymatic analysis of sixteen of its large or distinct RNAase T1-resistant oligonucleotides. Several WSN strain-specific nucleotide differences from the previously-determined sequence of NS1 mRNA from the PR8 (H0N1) strain of influenza A virus, were located within these sequences. The RNAase T1-resistant oligonucleotides were placed within the primary sequence of NS1 mRNA, using the PR8 strain sequence data. The resulting linear map was then used to identify NS2 mRNA isolated from the infected cell cytoplasm, and an NS-related RNA species generated from NS1 mRNA incubated in a HeLa cell-free extract.",

keywords = "Influenza mRNA, Primed RNA synthesis, RNA processing, mRNA",

author = "Nicholson, {Allen W.} and Frankfort, {Howard M.} and Davis, {Nicholas G.} and Stefano Ferrari and Lamb, {Robert A.} and Robertson, {Hugh D.}",

note = "Funding Information: Western Reserve University) for materials and assistance, Dr. Tamara Michaeli (Columbia University) for performing the Xenopus oocyte nuclear injections, and Dr. Andrea Branch for helpful advice in several aspects of this work. These investigations are supported in part by an NIH grant (GM 28294) to H.D.R. and an NIH postdoctoral fellowship (GM 07773) to A.W.N.",

year = "1986",

month = nov,

day = "13",

doi = "10.1016/0167-4781(86)90018-7",

language = "English (US)",

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pages = "153--163",

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T1 - Direct characterization of influenza viral NS1 mRNA and related sequences from infected HeLa cells and a cell-free transcription system

AU - Nicholson, Allen W.

AU - Frankfort, Howard M.

AU - Davis, Nicholas G.

AU - Ferrari, Stefano

AU - Lamb, Robert A.

AU - Robertson, Hugh D.

N1 - Funding Information: Western Reserve University) for materials and assistance, Dr. Tamara Michaeli (Columbia University) for performing the Xenopus oocyte nuclear injections, and Dr. Andrea Branch for helpful advice in several aspects of this work. These investigations are supported in part by an NIH grant (GM 28294) to H.D.R. and an NIH postdoctoral fellowship (GM 07773) to A.W.N.

PY - 1986/11/13

Y1 - 1986/11/13

N2 - The NS1 mRNA of the influenza A virus WSN (H0N1) strain was isolated from a cell-free transcription system, and from the cytoplasm of virus-infected HeLa cells. The 32P-labeled NS1 mRNA derived from the infected cell cytoplasm was characterized by the secondary enzymatic analysis of sixteen of its large or distinct RNAase T1-resistant oligonucleotides. Several WSN strain-specific nucleotide differences from the previously-determined sequence of NS1 mRNA from the PR8 (H0N1) strain of influenza A virus, were located within these sequences. The RNAase T1-resistant oligonucleotides were placed within the primary sequence of NS1 mRNA, using the PR8 strain sequence data. The resulting linear map was then used to identify NS2 mRNA isolated from the infected cell cytoplasm, and an NS-related RNA species generated from NS1 mRNA incubated in a HeLa cell-free extract.

AB - The NS1 mRNA of the influenza A virus WSN (H0N1) strain was isolated from a cell-free transcription system, and from the cytoplasm of virus-infected HeLa cells. The 32P-labeled NS1 mRNA derived from the infected cell cytoplasm was characterized by the secondary enzymatic analysis of sixteen of its large or distinct RNAase T1-resistant oligonucleotides. Several WSN strain-specific nucleotide differences from the previously-determined sequence of NS1 mRNA from the PR8 (H0N1) strain of influenza A virus, were located within these sequences. The RNAase T1-resistant oligonucleotides were placed within the primary sequence of NS1 mRNA, using the PR8 strain sequence data. The resulting linear map was then used to identify NS2 mRNA isolated from the infected cell cytoplasm, and an NS-related RNA species generated from NS1 mRNA incubated in a HeLa cell-free extract.

KW - Influenza mRNA

KW - Primed RNA synthesis

KW - RNA processing

KW - mRNA

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