Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2002 Dec;76(24):12951–12962. doi: 10.1128/JVI.76.24.12951-12962.2002

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2002, American Society for Microbiology

PMC Copyright notice

FIG. 2. — WSN-NS1(1-126)PUT3 and WSN-NS1(1-126)DmNcd viruses. (A) Schematic representation of the dimeric structures of the wild-type and mutant NS1(1-126), NS1(1-126)DmNcd, and NS1(1-126)PUT3 proteins. The crystal structure of the first 73 amino acids (aa) of the NS1 protein is adapted from reference 25. The crystal structure of the dimerization domain of DmNcd is adapted from reference 49. The structure of the dimerization domain of PUT3 is adapted from reference 50. The unknown structures are indicated by boxes. (B) Schematic representation of the NS genes and gene transcripts for influenza A WSN-NS1(1-126) dimerization domain-containing viruses, WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3. NS genes and mRNA transcripts of the top two viruses are schematically represented as in Fig. 1. The 6-glycine linker region in WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3 viruses is indicated by the striped box. The 24-amino-acid-long and 28-amino-acid-long dimerization domains from the D. melanogaster DmNcd protein or the S. cerevisiae PUT3 protein are represented by the checkered boxes. NEP, nuclear export protein; aa, amino acid; nt, nucleotide. (C) Western blot analysis of wild-type and mutant NS1 proteins in infected cells. MDBK cells were mock infected (mock) or were infected with wild-type (wt) WSN virus, recombinant WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus at an MOI of 1. Sixteen hours postinfection, cell extracts were made and subjected to Western blot analysis with antibodies against the viral NP and NS1 proteins. (D) Pulse-chase analysis of wild-type NS1 and mutant NS1 proteins. MDBK cells were either mock infected or infected with wild-type WSN virus, WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus at an MOI of 2. Four hours postinfection, cells were starved for methionine and cysteine and then pulse-labeled with ³⁵S-Met and ³⁵S-Cys for 30 min. At 1, 2, 4, and 20 h postchase total cell extracts were immunoprecipitated with antibodies against influenza NP and NS1 proteins. Samples were then boiled in SDS loading buffer and were separated by SDS-12% PAGE. The gel was then dried and subjected to autoradiography. h.p.c., hours postchase. (E) Pathogenicity of WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3 viruses in mice. Groups of five mice were intranasally infected with 2 × 10⁴ PFU of either wild-type WSN, WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus/mouse. Animals were inspected daily following infection. The percentage of surviving animals as a function of time is represented.

FIG. 2. — WSN-NS1(1-126)PUT3 and WSN-NS1(1-126)DmNcd viruses. (A) Schematic representation of the dimeric structures of the wild-type and mutant NS1(1-126), NS1(1-126)DmNcd, and NS1(1-126)PUT3 proteins. The crystal structure of the first 73 amino acids (aa) of the NS1 protein is adapted from reference 25. The crystal structure of the dimerization domain of DmNcd is adapted from reference 49. The structure of the dimerization domain of PUT3 is adapted from reference 50. The unknown structures are indicated by boxes. (B) Schematic representation of the NS genes and gene transcripts for influenza A WSN-NS1(1-126) dimerization domain-containing viruses, WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3. NS genes and mRNA transcripts of the top two viruses are schematically represented as in Fig. 1. The 6-glycine linker region in WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3 viruses is indicated by the striped box. The 24-amino-acid-long and 28-amino-acid-long dimerization domains from the D. melanogaster DmNcd protein or the S. cerevisiae PUT3 protein are represented by the checkered boxes. NEP, nuclear export protein; aa, amino acid; nt, nucleotide. (C) Western blot analysis of wild-type and mutant NS1 proteins in infected cells. MDBK cells were mock infected (mock) or were infected with wild-type (wt) WSN virus, recombinant WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus at an MOI of 1. Sixteen hours postinfection, cell extracts were made and subjected to Western blot analysis with antibodies against the viral NP and NS1 proteins. (D) Pulse-chase analysis of wild-type NS1 and mutant NS1 proteins. MDBK cells were either mock infected or infected with wild-type WSN virus, WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus at an MOI of 2. Four hours postinfection, cells were starved for methionine and cysteine and then pulse-labeled with ³⁵S-Met and ³⁵S-Cys for 30 min. At 1, 2, 4, and 20 h postchase total cell extracts were immunoprecipitated with antibodies against influenza NP and NS1 proteins. Samples were then boiled in SDS loading buffer and were separated by SDS-12% PAGE. The gel was then dried and subjected to autoradiography. h.p.c., hours postchase. (E) Pathogenicity of WSN-NS1(1-126)DmNcd and WSN-NS1(1-126)PUT3 viruses in mice. Groups of five mice were intranasally infected with 2 × 10⁴ PFU of either wild-type WSN, WSN-NS1(1-126), WSN-NS1(1-126)DmNcd, or WSN-NS1(1-126)PUT3 virus/mouse. Animals were inspected daily following infection. The percentage of surviving animals as a function of time is represented.