The B cell-specific nuclear factor OTF-2 positively regulates transcription of the human class II transplantation gene, DRA.
The promoter of the major histocompatibility class II gene DRA contains an octamer element (ATTTGCAT) that is required for efficient DRA expression in B cells. Several DNA-binding proteins are known to bind this sequence. The best characterized are the B cell-specific OTF-2 and the ubiquitous OTF-1. This report directly demonstrates that OTF-2 but not OTF-1 regulates the DRA gene. In vitro transcription analysis using protein fractions enriched for the octamer-binding protein OTF-2 demonstrate a positive functional role for OTF-2 in DRA gene transcription. In contrast, OTF-1-enriched protein fractions did not affect DRA gene transcription although it functionally enhanced the transcription of another gene. Recombinant OTF-2 protein produced by in vitro transcription/translation could also enhance DRA gene transcription in vitro. In vivo transient transfection studies utilizing an OTF-2 expression vector resulted in similar findings: that OTF-2 protein enhanced DRA gene transcription, and that this effect requires an intact octamer element. Together these results constitute the first direct evidence of a positive role for the lymphoid-specific octamer-binding factor in DRA gene transcription.
Zeleznik-Le
N J
NJ
Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill 27599-7295.
Itoh-Lindstrom
Y
Y
Clarke
J B
JB
Moore
T L
TL
Ting
J P
JP
D001345:Autoradiography; D001402:B-Lymphocytes / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D002855:Chromatography, Thin Layer; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D005802:Genes, MHC Class II; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D050813:Octamer Transcription Factor-2; D010957:Plasmids; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
D004268:DNA-Binding Proteins; D050813:Octamer Transcription Factor-2; C058435:POU2F2 protein, human; D011994:Recombinant Proteins; D014157:Transcription Factors; D004247:DNA