1887

Abstract

Summary

A mouse interferon alpha gene (MuIFN-α10) was isolated from a BALB/c cosmid genomic library. The gene was located on a 1.8 kb dIII fragment and a 5.1 kb RI fragment. The coding region and parts of the 5′ and 3′ non-coding regions were sequenced. The results showed that the MuIFN-α10 gene encoded a protein of 167 amino acids. Like most other MuIFN-α species it contained a putative -glycosylation site at amino acid positions 78 to 80. It also possessed cysteine residues at positions 1, 29, 86, 99 and 129. In the signal peptide, in addition to cysteine 21, which is present in all MuIFN-α species sequenced so far, a cysteine was found at position 22. At the amino acid level MuIFN-α10 showed strong homology to MuIFN-α1 (only 15 out of 167 amino acids were different). The MuIFN-α10 gene was transiently expressed in monkey COS cells under the direction of the simian virus 40 early promoter. The protein product secreted by COS cells was equally active on mouse (L929) and hamster (CHO) cells. However, as compared to MuIFN-α1 and MuIFN-α4 the specific activity on mouse cells of the protein was 10- to 100-fold lower. To find out which region of its structure was responsible for this low activity, hybrids of the genes encoding MuIFN-α10 and MuIFN-α1 were constructed using the two common I sites which correspond to positions between amino acids 67 and 68 and 123 and 124, respectively. The data showed that hybrid constructs which were MuIFN-α1-like from amino acid 68 or MuIFN-α10-like from position 124 to the C terminus possessed high antiviral activity. Other hybrid constructs were hardly active at all. This implied that the amino acid 68 to 123 region was mainly responsible for the low antiviral activity of MuIFN-α10. In this part of the molecule MuIFN-α1 and MuIFN-α10 differed in only five amino acids. A serine at position 110 and a valine at 85 were unique to MuIFN-α10 as compared to all known MuIFN-α and human IFN-α subspecies.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-69-1-67
1988-01-01
2019-08-19
Loading full text...

Full text loading...

http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-69-1-67
Loading

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error