Nucleic acids encoding a novel influenza virus non-structural protein (NS1)-binding host factor designated NS1I-1
View Patent ↗The present invention relates to the identification of host cell proteins that interact with viral proteins required for virus replication, and high throughput assays to identify compounds that interfere with the specific interaction between the viral and host cell protein. Interfering compounds that inhibit viral replication can be used therapeutically to treat viral infection. The invention is based, in part, on the Applicants' discovery of novel interactions between proteins of the influenza virus and a human host cell proteins. One of these host cell proteins, referred to herein as NPI-1, interacts with influenza virus protein NP, and may be an accessory protein required for replication of influenza virus. Another of these host cell proteins, referred to herein as NS1I-1, interacts with influenza virus protein NS 1 . Compounds that interfere with the binding of the host cell and viral proteins, and inhibit viral replication can be useful for treating viral infection in vivo.
1. An isolated nucleic acid comprising the nucleotide sequence of SEQ ID NO: 19, or the complement thereof.
2. An isolated nucleic acid comprising a nucleotide sequence which encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:20, or the complement thereof.
3. An isolated nucleic acid which hybridizes over its full length to the complement of the nucleotide sequence consisting of SEQ ID NO: 19 under highly stringent conditions comprising washing in 0.1×SSC/0.1% SDS at 68° C.
4. The isolated nucleic acid of claim 3 , which encodes a polypeptide that binds to influenza virus NS1.
5. An isolated nucleic acid comprising a nucleotide sequence which encodes a fusion polypeptide comprising the amino acid sequence encoded by the nucleic acid of claim 3 and a heterologous protein.
6. An isolated nucleic acid comprising a nucleotide sequence which encodes a fusion polypeptide comprising the amino acid sequence of SEQ ID NO:20 and a heterologous protein.
7. An expression vector comprising the nucleic acid of claim 1 operatively associated with a regulatory element that directs the expression of the nucleic acid.
8. An expression vector comprising the nucleic acid of claim 2 operatively associated with a regulatory element that directs the expression of the nucleic acid.
9. An expression vector comprising the nucleic acid of claim 3 operatively associated with a regulatory element that directs the expression of the nucleic acid.
10. An expression vector comprising the nucleic acid of claim 5 operatively associated with a regulatory element that directs the expression of the nucleic acid.
11. An expression vector comprising the nucleic acid of claim 6 operatively associated with a regulatory element that directs the expression of the nucleic acid.
12. A genetically engineered host cell comprising the nucleic acid of claim 1 operatively associated with a regulatory element that directs the expression of the nucleic acid.
13. A genetically engineered host cell comprising the nucleic acid of claim 2 operatively associated with a regulatory element that directs the expression of the nucleic acid.
14. A genetically engineered host cell comprising the nucleic acid of claim 3 operatively associated with a regulatory element that directs the expression of the nucleic acid.
15. A genetically engineered host cell comprising the nucleic acid of claim 5 operatively associated with a regulatory element that directs the expression of the nucleic acid.
16. A genetically engineered host cell comprising the nucleic acid of claim 6 operatively associated with a regulatory element that directs the expression of the nucleic acid.
17. A method for producing a polypeptide comprising: (a) culturing the host cell of claim 12 under conditions in which the nucleic acid is expressed, and (b) recovering the polypeptide produced.
18. A method for producing a polypeptide comprising: culturing the host cell of claim 13 under conditions in which the nucleic acid is expressed and (b) recovering the polypeptide produced.
19. A method for producing a polypeptide comprising: (a) culturing the host cell of claim 14 under conditions in which the nucleic acid is expressed, and (b) recovering the polypeptide produced.
20. A method for producing a polypeptide comprising: (a) culturing the host cell of claim 16 under conditions in which the nucleic acid is expressed, and (b) recovering the polypeptide produced.
21. A method for producing a polypeptide comprising: (a) culturing the host cell of claim 16 under conditions in which the nucleic acid is expressed, and (b) recovering the polypeptide produced.