Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells

W. Northemann, T. A. Braciak, Masahira Hattori, F. Lee, G. H. Fey

Research output: Contribution to journalArticle

168 Citations (Scopus)

Abstract

Rat peritoneal exudate cells produce two interleukin 6 (IL6) messenger RNA species, a major 1200 nucleotide and a 5-fold less abundant, 2400-nucleotide species. A cDNA clone representing the major species was isolated, and sequenced. The 1055-base pair insert covered the 3'-nontranslated region, the 211 triplet coding region and most of the 5'-nontranslated region. The derived rat IL6 amino acid sequence was 93 and 58% identical, respectively, with mature murine and human IL6. Rat IL6 lacks N-glycosylation sites but contains a fifth cysteinyl residue in addition to the 4 residues shared in conserved positions with murine and human IL6. Stable murine L cell and human HeLa-derived cell lines were established by cotransfection with rat IL6 cDNA and a selectable neomycin resistance marker. These lines secrete 9-fold increased amounts of functional IL6 over their respective parental cells. A stable rat macrophage-derived cell line, RM-SV1, was established by transformation with simian virus 40. IL6 and IL1 mRNA levels are inducible 20- and 3.5-fold, respectively, in this line by treatment with lipopolysaccharide with kinetics characteristic of macrophages. A set of three overlapping genomic DNA clones was isolated and a 10-kilobase DNA segment was sequenced containing the rat IL6 gene plus 2.9 kilobases of 5'-flanking and 1.3 kilobases of 3'-flanking sequences. The two transcription start sites used in RM-SV1 cells were mapped within 5 base pairs of each other. The exon/intron boundaries are conserved with the murine and human IL6 genes. The two IL6 mRNA species are generated by alternative polyadenylation at sites separated by a distance of 1.2 kilobases. The intervening region contains a repetitive element 72-80% identical with the rat and murine consensus L1 family sequences.

Original languageEnglish
Pages (from-to)16072-16082
Number of pages11
JournalJournal of Biological Chemistry
Volume264
Issue number27
Publication statusPublished - 1989
Externally publishedYes

Fingerprint

Macrophages
Rats
Interleukin-6
Genes
Gene Expression
Base Pairing
Messenger RNA
Nucleotides
Complementary DNA
Clone Cells
Cells
3' Flanking Region
Glycosylation
Cell Line
Polyadenylation
Simian virus 40
Neomycin
Transcription Initiation Site
DNA
Exudates and Transudates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Northemann, W., Braciak, T. A., Hattori, M., Lee, F., & Fey, G. H. (1989). Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells. Journal of Biological Chemistry, 264(27), 16072-16082.

Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells. / Northemann, W.; Braciak, T. A.; Hattori, Masahira; Lee, F.; Fey, G. H.

In: Journal of Biological Chemistry, Vol. 264, No. 27, 1989, p. 16072-16082.

Research output: Contribution to journalArticle

Northemann, W, Braciak, TA, Hattori, M, Lee, F & Fey, GH 1989, 'Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells', Journal of Biological Chemistry, vol. 264, no. 27, pp. 16072-16082.
Northemann, W. ; Braciak, T. A. ; Hattori, Masahira ; Lee, F. ; Fey, G. H. / Structure of the rat interleukin 6 gene and its expression in macrophage-derived cells. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 27. pp. 16072-16082.
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