Transcription factors Stat3 and Stat5b are present in rat liver nuclei late in an acute phase response and bind...

Proteins binding at the interleukin-6 response element of the rat alpha 2 macroglobulin gene were purified by a combination of chromatographic procedures including binding site-specific DNA-affinity chromatography as the principal step. Three polypeptides of 92, 91, and 86 kDa were enriched approximately 6,300-fold from nuclei of rat livers excised 12 h after the induction of an experimental acute phase response. Amino acid sequence analysis identified the 86- and 91-kDa species as two forms of the transcription factor Stat3 and the 92-kDa species as the factor Stat5b. This identification was confirmed by gel mobility shift-supershift experiments using specific antisera for Stat3 and Stat5. Unexpectedly, activated Stat5 was also detected in the nuclei of untreated control rats. cDNA clones representing Stat3 and two isoforms of Stat5b were isolated from a cDNA library prepared with mRNA from rat livers excised at the peak of an experimental acute phase response. Full-length Stat5b, predicted from cDNA, consisted of 786 amino acids, while the variant Stat5b delta 40C lacked 41 amino acids at the COOH terminus. The amino acid sequence of rat Stat5b showed 26.7% overall identity with rat Stat3, 87.3% with sheep Stat5a, 92.5% with murine Stat5a, and 98.7% with murine Stat5b.