A transcription reinitiation intermediate that is stabilized by activator

Abstract

High levels of gene transcription by RNA polymerase II depend on high rates of transcription initiation and reinitiation. Initiation requires recruitment of the complete transcription machinery to a promoter, a process facilitated by activators and chromatin remodelling factors. Reinitiation probably occurs through a different pathway1. After initiation, a subset of the transcription machinery remains at the promoter, forming a platform for assembly of a second transcription complex2,3,4. Here we describe the isolation of a reinitiation intermediate that includes transcription factors TFIID, TFIIA, TFIIH, TFIIE and Mediator. This intermediate can act as a scaffold for formation of a functional reinitiation complex. Formation of this scaffold is dependent on ATP and TFIIH. The scaffold is stabilized in the presence of the activator Gal4–VP16, but not Gal4–AH, suggesting a new role for some activators and Mediator in promoting high levels of transcription.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Scaffold contains activator, TFIID, TFIIA, Mediator, TFIIH and TFIIE.
Figure 2: Scaffold supports reinitiation.
Figure 3: Scaffold formation is TFIIH-dependent.
Figure 4: Gal4–VP16 promotes scaffold stability and a higher rate of reinitiation.
Figure 5: Reinitiation model.

References

  1. 1

    Hahn, S. Activation and the role of reinitiation in the control of transcription by RNA polymerase II. Cold Spring Harb. Symp. Quant. Biol. 63, 181–188 (1998).

    CAS  Article  Google Scholar 

  2. 2

    Roberts, S. G., Choy, B., Walker, S. S., Lin, Y. S. & Green, M. R. A role for activator-mediated TFIIB recruitment in diverse aspects of transcriptional regulation. Curr. Biol. 5, 508–516 (1995).

    CAS  Article  Google Scholar 

  3. 3

    Zawel, L., Kuman, K. P. & Reinberg, D. Recycling of the general transcription factors during RNA polymerase II transcription. Genes Dev. 9, 1479–1490 (1995).

    CAS  Article  Google Scholar 

  4. 4

    Sandaltzopoulos, R. & Becker, P. B. Heat shock factor increases the reinitiation rate from potentiated chromatin templates. Mol. Cell. Biol. 18, 361–367 (1998).

    CAS  Article  Google Scholar 

  5. 5

    Ranish, J. A., Yudkovsky, N. & Hahn, S. Intermediates in formation and activity of the RNA polymerase II preinitiation complex: holoenzyme recruitment and a postrecruitment role for the TATA box and TFIIB. Genes Dev. 13, 49–63 (1999).

    CAS  Article  Google Scholar 

  6. 6

    Kim, Y. J., Bjorklund, S., Li, Y., Sayre, M. H. & Kornberg, R. D. A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA Polymerase II. Cell 77, 599–608 (1994).

    CAS  Article  Google Scholar 

  7. 7

    Koleske, A. J. & Young, R. A. An RNA polymerase II holoenzyme responsive to activators. Nature 368, 466 –469 (1994).

    ADS  CAS  Article  Google Scholar 

  8. 8

    Jiang, Y. & Gralla, J. D. Uncoupling of initiation and reinitiation rates during HeLa RNA polymerase II transcription in vitro. Mol. Cell. Biol. 13, 4572–4577 (1993).

    CAS  Article  Google Scholar 

  9. 9

    Svejstrup, J. Q. et al. Evidence for a mediator cycle at the initiation of transcription. Proc. Natl Acad. Sci. USA 94, 6075– 6078 (1997).

    ADS  CAS  Article  Google Scholar 

  10. 10

    Kraus, W. L. & Kadonaga, J. T. p300 and estrogen receptor cooperatively activate transcription via differential enhancement of initiation and reinitiation. Genes Dev. 12, 331–342 (1998).

    CAS  Article  Google Scholar 

  11. 11

    Sheridan, P. L., Mayall, T. P., Verdin, E. & Jones, K. A. Histone acetyltransferases regulate HIV-1 enhancer activity in vitro. Genes Dev. 11, 3327–3340 (1997).

    CAS  Article  Google Scholar 

  12. 12

    Ho, S. N., Biggar, S. R., Spencer, D. M., Schreiber, S. L. & Crabtree, G. R. Dimeric ligands define a role for transcriptional activation domains in reinitiation. Nature 382, 822–826 ( 1996).

    ADS  CAS  Article  Google Scholar 

  13. 13

    Stringer, K. F., Ingles, C. J. & Greenblatt, J. Direct and selective binding of an acidic transcriptional activation domain to the TATA-box factor TFIID. Nature 345, 783–786 (1990).

    ADS  CAS  Article  Google Scholar 

  14. 14

    Ozer, J. et al. Molecular cloning of the small (γ) subunit of human TFIIA reveals functions critical for activated transcription. Genes Dev. 8, 2324–2335 ( 1994).

    CAS  Article  Google Scholar 

  15. 15

    Koh, S. S., Ansari, A. Z., Ptashne, M. & Young, R. A. An activator target in the RNA polymerase II holoenzyme. Mol. Cell 1, 895–904 ( 1998).

    CAS  Article  Google Scholar 

  16. 16

    Lee, Y. C., Park, J. M., Min, S., Han, S. J. & Kim, Y. J. An activator binding module of yeast RNA polymerase II holoenzyme. Mol. Cell. Biol. 19, 2967– 2976 (1999).

    CAS  Article  Google Scholar 

  17. 17

    Yean, D. & Gralla, J. Transcription reinitiation rate: a special role for the TATA box. Mol. Cell. Biol. 17 , 3809–3816 (1997).

    CAS  Article  Google Scholar 

  18. 18

    Klein, C. & Struhl, K. Increased recruitment of TATA-binding protein to the promoter by transcriptional activation domains in vivo. Science 266, 280–282 ( 1994).

    ADS  CAS  Article  Google Scholar 

  19. 19

    Xiao, H., Friesen, J. D. & Lis, J. T. Recruiting TATA-binding protein to a promoter: transcription activation without an upstream activator. Mol. Cell. Biol. 15, 5757–5761 (1995).

    CAS  Article  Google Scholar 

  20. 20

    Farrell, S., Simkovich, N., Wu, Y., Barberis, A. & Ptashne, M. Gene activation by recruitment of the RNA polymerase II holoenzyme. Genes Dev. 10, 2359– 2367 (1996).

    CAS  Article  Google Scholar 

  21. 21

    Keaveney, M. & Struhl, K. Activator-mediated recruitment of the RNA polymerase II machinery is the predominant mechanism for transcriptional activation in yeast. Mol. Cell 1, 917– 924 (1998).

    CAS  Article  Google Scholar 

  22. 22

    Hawley, D. K. & Roeder, R. G. Functional steps in transcription initiation and reinitiation from the major late promoter in a HeLa nuclear extract. J. Biol. Chem. 262, 3452– 3461 (1987).

    CAS  PubMed  Google Scholar 

  23. 23

    Holstege, F. C. et al. Dissecting the regulatory circuitry of a eukaryotic genome. Cell 95, 717–728 (1998).

    CAS  Article  Google Scholar 

  24. 24

    Guzder, S. N. et al. DNA repair gene RAD3 of S. cerevisiae is essential for transcription by RNA polymerase II. Nature 367, 91– 94 (1994).

    ADS  CAS  Article  Google Scholar 

  25. 25

    Cismowski, M. J., Laff, G. M., Solomon, M. J. & Reed, S. I. KIN28 encodes a C-terminal domain kinase that controls mRNA transcription in S. cerevisiae but lacks cyclin-dependent kinase-activating kinase (CAK) activity. Mol. Cell. Biol. 15, 2983– 2992 (1995).

    CAS  Article  Google Scholar 

  26. 26

    Kang, J. J., Auble, D. T., Ranish, J. A. & Hahn, S. Analysis of the yeast transcription factor TFIIA: distinct functional regions and a polymerase II-specific role in basal and activated transcription. Mol Cell Biol 15, 1234-1243 ( 1995).

    Article  Google Scholar 

  27. 27

    Ranish, J. A. & Hahn, S. The yeast general transcription factor TFIIA is composed of two polypeptide subunits. J. Biol. Chem. 266, 19320–19327 (1991).

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank members of the Hahn and Reeder laboratories for helpful discussions, and A. Krumm, S. Parkhurst and R. Reeder for comments on the manuscript. We also thank L. Prakash for providing the Rad3ts strain, M. Solomon for providing the Kin28ts strain, D. Reinberg for TFIIE antibodies and H. Sakurai for providing the Tfa1ts strain and antibodies to Gal11. This work was supported by grants from the NIH to S.H. and an NIH training grant to N.Y. S.H. is an associate investigator of the Howard Hughes Medical Institute.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Steven Hahn.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Yudkovsky, N., Ranish, J. & Hahn, S. A transcription reinitiation intermediate that is stabilized by activator . Nature 408, 225–229 (2000). https://doi.org/10.1038/35041603

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing