The DNA binding domain (POU domain) of transcription factor oct-1 suffices for stimulation of DNA replication.
Oct-1, also referred to as NFIII, OTF-1, OBP100 or NF-A1, is a ubiquitous sequence-specific DNA binding protein that activates transcription and adenovirus DNA replication. The protein contains a conserved DNA binding domain (POU domain) present in several transcription factors. We have overproduced oct-1, the related oct-2 and several oct-1 deletion mutants in a vaccinia expression system to identify the domains important for activation of DNA replication in vitro. Both oct-1 and oct-2 stimulate adenovirus DNA replication in an octamer-dependent manner. From deletion studies it appears that the 160 amino acid long POU domain suffices for stimulation. This domain consists of two subdomains, a POU-specific and a homeo domain. Deletion of the POU-specific domain revealed that the homeo domain has an intrinsic, but weak DNA binding activity and surprisingly, inhibits DNA replication. As the POU domain does not coincide with the transcription activation domain, these results indicate that, although oct-1 functions both in DNA replication and transcription, the mechanisms underlying these processes are probably distinct.
Verrijzer
C P
CP
Laboratory for Physiological Chemistry, University of Utrecht, The Netherlands.
Kal
A J
AJ
Van der Vliet
P C
PC
D000260:Adenoviruses, Human / Q000235:genetics* / Q000254:growth & development; D001483:Base Sequence; D004261:DNA Replication; D004274:DNA, Recombinant / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I; D006367:HeLa Cells; D051863:Host Cell Factor C1; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D050811:Octamer Transcription Factor-1; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication
D004274:DNA, Recombinant; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D051863:Host Cell Factor C1; D050811:Octamer Transcription Factor-1; C494165:POU2F1 protein, human; D014157:Transcription Factors; D003850:Deoxyribonuclease I