High mobility group proteins 1 and 2 function as general class II transcription factors

Abstract

High mobility group (HMG) proteins 1 and 2 are thought to be associated with chromatin enriched in active gene sequences, to stimulate endogenous transcription of class II and III genes using HMG-depleted nuclei, and to bind specific DNA sequences upstream of the coding regions of trout HMG-T and human beta-globin genes. In testing the possibility that these proteins may act as general transcription factors, the run-off transcription of trout protamine, human beta-globin, adenovirus 2 major late promoter, and herpes simplex virus (HSV) thymidine kinase genes was found to be inhibited by affinity-purified HMG-1 and -2 antibodies. The inhibition was partially relieved by exogenously added HMG-1 or -2. A complementation assay showed that the 0.15 M KCl flowthrough of HeLa nuclear extract fractionated by anion-exchange chromatography (DE-52) could be replaced by purified HMG-1 and/or -2 to complement transcription of the trout protamine gene by the 0.5 M KCl eluate fraction. Inhibition studies with heparin showed that HMG-1 and -2 were required for initiation of transcription. These results indicate an absolute requirement of HMG-1 and -2 for class II gene transcription. Western blotting and transcription reconstituted with purified factors show a copurification of HMG-1 and -2 with factor II B, described earlier by Reinberg and Roeder [(1987) J. Biol. Chem. 262, 3310-3321].