NGFIA (EGR1) contains transcription activating domains in both the amino terminal and carboxyl terminal regions of the protein.
We constructed and tested a number of lac repressor fusion proteins containing various portions of the zinc-finger containing protein NGFIA for their ability to stimulate transcription of a reporter gene containing lac operators. NGFIA contains two transcription activation regions, found in two distinct regions of the protein. The carboxyl (C) terminal portion of the molecule contains a weak activation domain, including five tandem copies of an eight amino acid repeat (T/S,T/S,F/Y,P,S,P,X,X). These five tandem copies of the repeated sequence activated reporter gene transcription 4-7 fold. Amino acids 1 through 293 in the amino (N) terminus of NGFIA function as a strong transcription activation domain stimulating transcription up to 80-fold. Fusions including amino acids 1-393 failed to activate transcription, indicating the presence of a domain capable of suppressing the N-terminal transcriptional activation function.
Division of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115.
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C
DeFranco
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Ro
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Grossel
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English
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Hansen
J A
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Wagner
J D
JD
Licht
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007763:Lac Operon; D008969:Molecular Sequence Data; D016716:PC12 Cells; D010957:Plasmids; D011401:Promoter Regions, Genetic; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology
D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D004247:DNA; D013937:Thymidine Kinase