Activation of octamer-containing promoters by either octamer-binding transcription factor 1 (OTF-1) or OTF-2 and requirement of an additional B-cell-specific component for optimal transcription of immunoglobulin promoters.
Several distinct octamer-binding transcription factors (OTFs) interact with the sequence ATTTGCAT (the octamer motif), which acts as a transcription regulatory element for a variety of differentially controlled genes. The ubiquitous OTF-1 plays a role in expression of the cell cycle-regulated histone H2b gene as well as several other genes, while the tissue-specific OTF-2 has been implicated in the tissue-specific expression of immunoglobulin genes. In an attempt to understand the apparent transcriptional selectivity of these factors, we have investigated the physical and functional characteristics of OTF-1 purified from HeLa cells and both OTF-1 and OTF-2 purified from B cells. High-resolution footprinting and mobility shift-competition assays indicated that these factors were virtually indistinguishable in binding affinities and DNA-protein contacts on either the H2b or an immunoglobulin light-chain (kappa) promoter. In addition, each of the purified factors showed an equivalent intrinsic capacity to activate transcription from either immunoglobulin promoters (kappa and heavy chain) or the H2b promoter in OTF-depleted HeLa and B-cell extracts. However, with OTF-depleted HeLa extracts, neither factor could restore immunoglobulin gene transcription to the relatively high level observed in unfractionated B-cell extracts. Restoration of full immunoglobulin gene activity appears to require an additional B-cell regulatory component which interacts with the OTFs. The additional B-cell factor could act either by facilitating interaction of OTF activation domains with components of the general transcriptional machinery or by contributing a novel activation domain.
Pierani
A
A
Laboratory of Biochemistry and Molecular Biology, Rockefeller University, New York, New York 10021.
Heguy
A
A
Fujii
H
H
Roeder
R G
RG
D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003850:Deoxyribonuclease I; D005786:Gene Expression Regulation; D005803:Genes, Immunoglobulin; D006367:HeLa Cells / Q000276:immunology / Q000378:metabolism; D006657:Histones / Q000235:genetics; D051863:Host Cell Factor C1; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D050811:Octamer Transcription Factor-1; D050813:Octamer Transcription Factor-2; D015345:Oligonucleotide Probes; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D006657:Histones; D051863:Host Cell Factor C1; D007147:Immunoglobulin Light Chains; D050811:Octamer Transcription Factor-1; D050813:Octamer Transcription Factor-2; D015345:Oligonucleotide Probes; C494165:POU2F1 protein, human; C058435:POU2F2 protein, human; D014157:Transcription Factors; D003850:Deoxyribonuclease I