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Titolo:
CLEAVAGE OF THE NASCENT TRANSCRIPT INDUCED BY TFIIS IS INSUFFICIENT TO PROMOTE READ-THROUGH OF INTRINSIC BLOCKS TO ELONGATION BY RNA-POLYMERASE-II
Autore:
CIPRESPALACIN G; KANE CM;
Indirizzi:
UNIV CALIF BERKELEY,DEPT BIOCHEM & MOLEC BIOL,401 BARKER HALL BERKELEY CA 94720
Titolo Testata:
Proceedings of the National Academy of Sciences of the United Statesof America
fascicolo: 17, volume: 91, anno: 1994,
pagine: 8087 - 8091
SICI:
0027-8424(1994)91:17<8087:COTNTI>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
ASCITES TUMOR-CELLS; DEPENDENT TRANSCRIPTION; TERNARY COMPLEXES; ESCHERICHIA-COLI; DNA; PROTEIN; BINDING; GENE; TERMINATION; INVITRO;
Keywords:
GENE REGULATION; ELONGATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
36
Recensione:
Indirizzi per estratti:
Citazione:
G. Ciprespalacin e C.M. Kane, "CLEAVAGE OF THE NASCENT TRANSCRIPT INDUCED BY TFIIS IS INSUFFICIENT TO PROMOTE READ-THROUGH OF INTRINSIC BLOCKS TO ELONGATION BY RNA-POLYMERASE-II", Proceedings of the National Academy of Sciences of the United Statesof America, 91(17), 1994, pp. 8087-8091

Abstract

RNA polymerases encounter a variety of types of blocks to elongation during transcription in eukaryotic cells. At least one protein, TFIIS,can promote read-through of many types of blocks to elongation by RNApolymerase II, and this protein stimulates cleavage of the nascent transcript in stalled elongation complexes as a prelude to read-through. The C-terminal half of the TFIIS protein is sufficient for stimulating the cleavage and read-through reactions in vitro. To study how TFIISchanges the response of RNA polymerase II elongation complexes to such blocks, targeted amino acids in the C terminus of HeLa TFIIS were mutated to alanines. Two mutant TFIIS proteins as well as the unmutated C-terminal half of the TFIIS protein were purified following overexpression in Escherichia coli. Each protein was examined for read-through activity and ability to stimulate transcript cleavage in ternary elongation complexes. Mutant TFIIS5 (E174A, E175A) was reduced in read-through and cleavage activities relative to the unmutated, truncated TFIIS(Delta TFIIS). Mutant TFIIS7 (KI87A, K189A) was able to stimulate cleavage nearly at the rate and to the extent of the TFIIS5 mutant. In contrast to what was observed with TFIIS5, no detectable read-through was observed in the presence of the TFIIS7 mutant during the course of the reaction. Thus, there is no simple, direct correlation between the ability of TFIIS to promote cleavage and its ability to promote read-through by RNA polymerase II. These results suggest that although TFIISis necessary to mediate the cleavage reaction that precedes the read-through event, the cleavage event itself is not sufficient to allow read-through by RNA polymerase II.

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Documento generato il 01/10/20 alle ore 04:03:13