Regulation of TFIIH ATPase and kinase activities by TFIIE during active initiation complex formation

Abstract

The general transcription factor TFIIE, together with other general transcription factors, is essential for transcription initiation by RNA polymerase II. TFIIE stimulates the TFIIH-dependent kinase activity that phosphorylates the carboxy-terminal domain of the largest subunit of RNA polymerase II, and possesses a helicase activity. Here we show that human TFIIH has DNA-dependent ATPase activity and we characterize the stimulatory effect of TFIIE on both the ATPase and kinase activities. We demonstrate that extensive phosphorylation of RNA polymerase II occurs in a TFIIE-dependent manner in both the absence and presence of DNA but, in the latter case, only at a late stage of preinitiation complex assembly. We also show that TFIIH specifically phosphorylates three general transcription factors, human TFIID tau (TBP), TFIIE-alpha and TFIIF-alpha (RAP74).

Modulation of TFIIH‐associated kinase activity by complex formation and its relationship with CTD phosphorylation of RNA polymerase II

“…3C, lanes 5±7) although TFIIH strongly phosphorylated the CTD and caused a shift of the IIa form to the IIo form as previously reported (Fig. 3C, lanes 8±10 and Lu et al 1992;Ohkuma & Roeder 1994). As seen under solution conditions, TFIIE stimulated CTD phosphorylation under conditions of PIC formation (Fig.…”

“…As it has been demonstrated that TFIIH phosphorylates the CTD of PolII during the formation of the active PIC as well as in solution (Lu et al 1992;Ohkuma & Roeder 1994), we next examined CTD phosphorylation by CAK under the conditions of PIC formation (Fig. 3C).…”

Phosphorylation of the N‐terminal domain of Xenopus TATA‐box binding protein by DNA‐dependent protein kinase depends on the C‐terminal core domain

“…TBP was reported to be hyperphosphorylated during mitosis, but the protein kinase responsible for this modification has not been identified [11]. Furthermore, the TFIIH-associated kinase was found to phosphorylate the N-terminal domain of human TBP in vitro [12]. The present report adds to our knowledge of TBP phosphorylation by showing that Xenopus TBP (xTBP) is phos-*Corresponding author.…”

Regulation of RNA polymerase II activity by CTD phosphorylation and cell cycle control

“…These include the GTFs TBP, TFIIEa, TFIIFa (Ohkuma and Roeder, 1994;Yankulov and Bentley, 1997), Oct transcription factors (Inamoto et al, 1997), p53 (Ko et al, 1997), retinoic receptor alpha (RARa) (Rochette-Egly et al, 1997), estrogen receptor alpha (ERa) (Chen et al, 2000), and pRb (Wu et al, 2001). The speci®city of CDK7-cyclin H kinase activity towards various substrates appears to critically depend on its association with higher-order protein complexes.…”