Abstract

Paramyxoviruses are the major cause of respiratory disease in children. The long-term objective is to develop means to prevent respiratory disease caused by paramyxoviruses. To reach that objective, the proposed research will focus on structure-function relationships of the hemagglutinin-neuraminidase (HN) envelope glycoprotein of Newcastle disease virus (NDV), a prototypic paramyxovirus, in virus attachment, neuraminidase and fusion-promoting activity.
The aims outline Dr. Portner's strategy to achieve his goals.
Aim 1 : Proposal: To establish a high resolution three-dimensional (3-D) structure of HN.
Aim 2 : Proposal: To determine directly the location and 3-D structure of the receptor binding, neuraminidase, and fusion promoting activities of HN. Dr. Portner's laboratory has crystallized the NDV HN protein by the hanging drop-vapor diffusion method. These crystals diffract X-rays at moderately high resolution (up to 2.6A) indicative that they will be suitable for determination of the HN 3-D structure.
In Aim 1, the preparation of large amounts of larger crystals will be pursued with the prospect of increasing the resolution and to aid in obtaining and analyzing heavy metal derivatives essential to the solution of the HN structure.
Aim 2 will determine directly the location and spacial arrangement of regions tentatively defined as the receptor binding and neuraminidase (NA) sites by analyzing the 3-D structure of HN complexed with sialic acid analogues. Supporting evidence will come from mutational analysis of cell binding and NA activities expressed from cDNAs that contain site-directed mutations in the proposed NA and cell binding sites, and isolation and sequence analysis of receptor specificity mutations and escape mutants. Mutational analysis and knowledge of the 3-D structure should provide an assessment of the location and structure of the fusion promoting domain associated with the globular head region of HN, thus providing additional targets for drug intervention.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI038956-02
Application #
2633566
Study Section
Virology Study Section (VR)
Project Start
1997-01-01
Project End
1999-12-31
Budget Start
1998-01-01
Budget End
1998-12-31
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105

  Publications

Krishnamurthy, Sateesh; Takimoto, Toru; Scroggs, Ruth Ann et al. (2006) Differentially regulated interferon response determines the outcome of Newcastle disease virus infection in normal and tumor cell lines. J Virol 80:5145-55
Alymova, Irina V; Portner, Allen; Takimoto, Toru et al. (2005) The novel parainfluenza virus hemagglutinin-neuraminidase inhibitor BCX 2798 prevents lethal synergism between a paramyxovirus and Streptococcus pneumoniae. Antimicrob Agents Chemother 49:398-405
Zaitsev, Viatcheslav; von Itzstein, Mark; Groves, Darrin et al. (2004) Second sialic acid binding site in Newcastle disease virus hemagglutinin-neuraminidase: implications for fusion. J Virol 78:3733-41
Bousse, Tatiana L; Taylor, Garry; Krishnamurthy, Sateesh et al. (2004) Biological significance of the second receptor binding site of Newcastle disease virus hemagglutinin-neuraminidase protein. J Virol 78:13351-5
Takimoto, Toru; Portner, Allen (2004) Molecular mechanism of paramyxovirus budding. Virus Res 106:133-45
Connaris, Helen; Takimoto, Toru; Russell, Rupert et al. (2002) Probing the sialic acid binding site of the hemagglutinin-neuraminidase of Newcastle disease virus: identification of key amino acids involved in cell binding, catalysis, and fusion. J Virol 76:1816-24
Takimoto, Toru; Taylor, Garry L; Connaris, Helen C et al. (2002) Role of the hemagglutinin-neuraminidase protein in the mechanism of paramyxovirus-cell membrane fusion. J Virol 76:13028-33
Bousse, Tatiana; Matrosovich, Tatyana; Portner, Allen et al. (2002) The long noncoding region of the human parainfluenza virus type 1 f gene contributes to the read-through transcription at the m-f gene junction. J Virol 76:8244-51
Takimoto, T; Murti, K G; Bousse, T et al. (2001) Role of matrix and fusion proteins in budding of Sendai virus. J Virol 75:11384-91
Suzuki, T; Portner, A; Scroggs, R A et al. (2001) Receptor specificities of human respiroviruses. J Virol 75:4604-13

Showing the most recent 10 out of 13 publications