Identification and expression analysis of cDNAs encoding channel catfish type I interferons.
Previously a cDNA encoding a putative interferon gene, designated CF IFN-1, was identified from a catfish EST library. However, its constitutive expression, absence of a signal peptide, and apparently low level of biological activity suggested that this cDNA likely encoded an expressed pseudogene. Since Southern blot analysis suggested the presence of two to three IFN genes, additional cDNAs were generated from catfish fibroblast and lymphoid cell lines using primers designed to conserved regions of zebrafish and catfish interferon. Using this approach, three novel CF IFN genes, two of which likely encode functional interferon molecules, were identified. At the amino acid level, similarity among CF IFNs ranged from 71% to 82%, whereas similarity to other fish IFNs ranged from 15% to 35%. Although CF IFN-3, like CF IFN-1, lacks a signal peptide, CF IFN-2 and -4 appear to encode full-length, signal sequence-bearing genes. Consistent with their putative identification as functional genes, CF IFN-2 and -4 were not expressed in unstimulated cell lines, and CF IFN-2 was rapidly upregulated in CCO cells in response to virus infection or treatment with dsRNA. Moreover, as with salmon, fugu, and zebrafish interferon genes, CF IFN-1 contained four introns whose locations were conserved not only with respect to other fish IFNs, but also with respect to mammalian IFN-lambda. While it is likely that CF IFNs represent Type I IFNs, several characteristics preclude assigning these cytokines to any particular subfamily.
Long
Scott
S
Department of Microbiology, University of Mississippi Medical Center, 2500 North State Street, Jackson, 39216, USA.
Milev-Milovanovic
Ivanka
I
Wilson
Melanie
M
Bengten
Eva
E
Clem
L W
LW
Miller
N W
NW
Chinchar
V G
VG
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology; D005091:Exons; D020869:Gene Expression Profiling / Q000662:veterinary; D023061:Gene Order; D007059:Ictaluridae / Q000235:genetics* / Q000276:immunology*; D007370:Interferon Type I / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D007438:Introns; D008969:Molecular Sequence Data; D010802:Phylogeny; D012087:Reoviridae / Q000276:immunology / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D016415:Sequence Alignment / Q000662:veterinary; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D018076:DNA, Complementary; D007370:Interferon Type I