Complexes of yeast RNA polymerase II and RNA are substrates for TFIIS-induced RNA cleavage.
In the absence of DNA, purified yeast RNA polymerase II can bind RNA to form a binary complex. RNA in such RNA-RNA polymerase complexes undergoes reactions previously thought to be unique to nascent RNA in ternary complexes with DNA, including TFIIS-dependent cleavage and elongation by 3'-terminal addition of NMP from NTP. Both of these reactions are inhibited by alpha-amanitin. Hence, by several criteria the RNA in binary complexes is bound to the polymerase in a manner quite similar to that in ternary complexes in which the catalytic site for nucleotide addition is positioned at or near the 3'-OH terminus of the RNA. These findings are consistent with a model for the RNA polymerase ternary complex in which the RNA is bound at the 3' terminus through two protein-binding sites located up to 10 nt apart.
Johnson
T L
TL
Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720.
Chamberlin
M J
MJ
D000546:Amanitins / Q000494:pharmacology; D001483:Base Sequence; D003902:Detergents / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D008274:Magnesium / Q000378:metabolism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009711:Nucleotides / Q000378:metabolism; D011485:Protein Binding; D012319:RNA Polymerase II / Q000378:metabolism*; D012331:RNA, Fungal / Q000378:metabolism*; D012521:Sarcosine / Q000031:analogs & derivatives / Q000494:pharmacology; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism*; D035165:Transcription Factors, General; D014158:Transcription, Genetic; D035602:Transcriptional Elongation Factors
D000546:Amanitins; D003902:Detergents; D009711:Nucleotides; D012331:RNA, Fungal; D014157:Transcription Factors; D035165:Transcription Factors, General; D035602:Transcriptional Elongation Factors; C018648:transcription factor S-II; C025231:sarkosyl; D012319:RNA Polymerase II; D008274:Magnesium; D012521:Sarcosine