A Highly Purified RNA Polymerase II Elongation Control System

• Published in: The Journal of biological chemistry 2001-11, Vol.276 (45), p.42601-42609
• Main Authors: Renner, Dan B., Yamaguchi, Yuki, Wada, Tadashi, Handa, Hiroshi, Price, David H.
• Format: Article
• Language: English
• Subjects: 5,6-dichloro-1-^b-D-ribofuranosylbenzimidazole; DSIF protein; negative elongation factor; NELF protein; Nuclear Proteins - isolation & purification; Nuclear Proteins - physiology; P-TEFb protein; Positive Transcriptional Elongation Factor B; Protein-Serine-Threonine Kinases - physiology; RNA Polymerase II - physiology; TFIIF protein; Transcription Factors - isolation & purification; Transcription Factors - physiology; Transcription Factors, TFII; Transcription, Genetic; Transcriptional Elongation Factors
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M104967200

Studying the sensitivity of transcription to the nucleotide analog 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole has led to the discovery of a number of proteins involved in the regulation of transcription elongation by RNA polymerase II. We have developed a highly purified elongation control system composed of three purified proteins added back to isolated RNA polymerase II elongation complexes. Two of the proteins, 5,6-dichloro-1-β-d-ribofuranosylbenzimidazole sensitivity-inducing factor (DSIF) and negative elongation factor (NELF), act as negative transcription elongation factors by increasing the time the polymerase spent at pause sites. P-TEFb reverses the negative effect of DSIF and NELF through a mechanism dependent on its kinase activity. TFIIF is a general initiation factor that positively affects elongation by decreasing pausing. We show that TFIIF functionally competes with DSIF and NELF, and this competition is dependent on the relative concentrations of TFIIF and NELF.