Expression, Characterization, and Genomic Structure of Carp JAK1 Kinase Gene
Resource
DNA and Cell Biology 15 (10): 827-844
Journal
DNA and Cell Biology
Journal Volume
15
Journal Issue
10
Pages
827-844
Date Issued
1996
Date
1996
Author(s)
LEU, JIANN-HORNG
HUANG, FORE-LIEN
CHOU, CHEN-KUNG
HUANG, CHANG-JEN
LO, TUNG-BIN
Abstract
A 3.7-kb cDNA encodes the carp JAK1 kinase of 1,156 amino acid residues. The overall amino acid sequence identity between carp JAK1 and murine JAK1, JAK2, JAK3, and human TYK2 is 57%, 35.5%, 31.3%, and 42.4%, respectively. In addition, carp JAK1 shows higher sequence homology to mammalian JAK1 in both the kinase-like (JH2) and kinase (JH1) domains (approximately 70% identity). Therefore, carp JAK1 is a homolog of mammalian JAK1. To investigate the possible function of JH2 domain, full-length, and various truncated forms of carp JAK1 were produced in the baculovirus system. Our results demonstrate that c-JH1 and c-JH2 associate with each other and c-JH2 can be tyrosine- phosphorylated by c-JAK1 and by c-JH(1 + 2). The JAK1 gene was also isolated from a carp genomic library and characterized. This gene is divided into 24 exons spanning at least 31 kb of genomic DNA. Exon 1 contains the 5'- untranslated region and exon 2 contains the putative translation initiation site. The 2.5-kb DNA region upstream of the transcription initiation site contains numerous potential binding sites for transcription factors including NF-IL6, HNF-5, AP1, GHF-5, and E2A. When this DNA fragment was placed upstream of the chloramphenicol acetyltransferase (CAT) reporter gene and transfected into a carp CF cell line, it could drive the synthesis of CAT enzyme 16 times more efficiently than the promoterless pCAT-Basic. Deletion analysis defined a positive regulatory region between -1,023 and -528. A smaller region (-181 to +59) without any typical TATA-box sequences, G + C- rich sequences, or other binding sequences for known transcription factors still had promoter activity. Constructs without this region did not have detectable promoter activity. This suggests that this region of DNA may play an important role in the expression of carp JAK1 gene.
Other Subjects
phosphotransferase; amino acid sequence; animal cell; article; carp; exon; gene expression; gene structure; genetic transfection; nonhuman; priority journal; promoter region; reporter gene; sequence homology; TATA box; transcription initiation; Animalia; Cyprinus carpio; Felis catus; Mammalia; Murinae; unidentified baculovirus
Type
journal article
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