Stimulation of transcription in cultured cells by high mobility group protein 1: essential role of the acidic carboxyl-terminal region.
Several in vitro studies have suggested that high mobility group (HMG) protein 1 has a role in gene regulation as a trans activator or quasi-transcription factor. However, data on the molecular functions of HMG1 protein in these reactions are contradictory or obscure. In order to assess whether HMG1 protein does, in fact, have transcriptional activation potential, two assay systems in cultured cells were employed. HMG1 protein introduced into COS-1 cells as a complex with a reporter plasmid carrying the lacZ gene enhanced the level of the gene expression. Cotransfection of an expression plasmid carrying HMG1 cDNA into the cells with the reporter plasmid enhanced the activity of beta-galactosidase 2-3-fold in comparison with that of the control effector plasmid. The enhancement was proved to be dependent not on the replication but on the transcription of the reporter plasmid. In the cotransfection experiments, an expression plasmid the HMG1 molecule lacking the acidic carboxyl terminus repressed the expression of the reporter gene. The binding of an HMG1 protein variant lacking the acidic carboxyl terminus to DNA gave an extremely large shift of gel retardation in comparison with the complete HMG1 molecule. Together, these results indicate that HMG1 protein can enhance expression in cells in culture at the step of gene transcription and that the DNA binding domains comprising two-thirds of the HMG1 protein molecule are responsible for the inhibition property. Also, the acidic terminus of the HMG1 molecule is essential for the enhancement of gene expression in addition to elimination of the repression caused by the DNA binding. (ABSTRACT TRUNCATED AT 250 WORDS)
Aizawa
S
S
Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan.
Nishino
H
H
Saito
K
K
Kimura
K
K
Shirakawa
H
H
Yoshida
M
M
D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology*; D005822:Genetic Vectors; D006609:High Mobility Group Proteins / Q000737:chemistry* / Q000502:physiology*; D010957:Plasmids; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000737:chemistry* / Q000502:physiology*; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D014157:Transcription Factors; D001616:beta-Galactosidase