Site-directed mutagenesis of arginine 246, glutamic acid 247, and histidine 248 in the eukaryotic transcription factor S-II.
When His248 of the transcription factor S-II was replaced by alanine or tyrosine, the activity of the resulting mutants was less than 30% of that of the wild-type S-II. When Arg246, Glu247, and His248 were all replaced by leucine, the resulting mutant showed complete loss of activity. These results indicate that the amino acid sequence Arg-Glu-His at positions 246-248 of S-II is important for its stimulatory activity. The mutant S-II with no activity could not form a complex with RNA polymerase II, unlike wild-type S-II, but retained ability to interact with DNA.
Horikoshi
N
N
Faculty of Pharmaceutical Sciences, University of Tokyo, Japan.
Sekimizu
K
K
Natori
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D001483:Base Sequence; D002918:Chymotrypsin; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005971:Glutamates; D018698:Glutamic Acid; D006639:Histidine; D007700:Kinetics; D007930:Leucine; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D010446:Peptide Fragments / Q000302:isolation & purification; D010957:Plasmids; D012319:RNA Polymerase II / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors
D005971:Glutamates; D015345:Oligonucleotide Probes; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014157:Transcription Factors; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; D018698:Glutamic Acid; D006639:Histidine; D001120:Arginine; D012319:RNA Polymerase II; D002918:Chymotrypsin; D007930:Leucine