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Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II

Nature Cell Biology volume 6, pages 1094–1101 (2004)Cite this article

Abstract

Actin is abundant in the nucleus and has been implicated in transcription; however, the nature of this involvement has not been established. Here we demonstrate that β-actin is critically involved in transcription because antibodies directed against β-actin, but not muscle actin, inhibited transcription in vivo and in vitro. Chromatin immunoprecipitation assays demonstrated the recruitment of actin to the promoter region of the interferon-γ-inducible MHC2TA gene as well as the interferon-α-inducible G1P3 gene. Further investigation revealed that actin and RNA polymerase II co-localize in vivo and also co-purify. We employed an in vitro system with purified nuclear components to demonstrate that antibodies to β-actin block the initiation of transcription. This assay also demonstrates that β-actin stimulates transcription by RNA polymerase II. Finally, DNA-binding experiments established the presence of β-actin in pre-initiation complexes and also showed that the depletion of actin prevented the formation of pre-initiation complexes. Together, these data suggest a fundamental role for actin in the initiation of transcription by RNA polymerase II.

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Figure 1: Antibodies to β-actin inhibit RNA polymerase II transcription in vivo.
Figure 2: Antibodies to β-actin inhibit RNA polymerase II transcription in vitro.
Figure 3: β-actin is associated with RNA polymerase II.
Figure 4: Antibodies to β-actin and NEM-modified S1 inhibit transcription in a purified system.
Figure 5: Recruitment of actin to the MHC2TA promoter IV and the G1P3 promoter.
Figure 6: Antibodies to β-actin inhibit transcription initiation.
Figure 7: Actin is required for PIC formation.

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Acknowledgements

We thank P. Raychaudhury (Department of Biochemistry, University of Illinois at Chicago; UIC) for providing HeLa cells for purifying RNA polymerase II; M. Vigneron (Illkirch, France) for providing antibodies to RNA polymerase II; R. Moss (University of Wisconsin) for providing NEM-S1; M. L. Chen (Research Resources Center, UIC) for assistance with the confocal microscopy; and S. Huang (Northwestern University) for assistance with the in vivo transcription assay. V.F. was supported by the Grant Agency of the Czech Republic (Reg. No 304/03/P118). P.H. was supported by the Grant Agency of the Czech Republic (Reg. No. 304/01/0661 and 2004/04/0108), by the Grant Agency of the Academy of Sciences of the Czech Republic (Reg. No. IAA5039202) and by the Institutional Grant No. AV0Z5039906. Additional support was provided by grants from the US Public Health Service to P.H. (NSF/MSMT Program ME143), J.L.L. (HL57291), J.A.G. (GM55235), T.T.H. (AI 047770) and P.de.L. (NSF INT 9724168 and NIH HL 56489).

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Authors and Affiliations

  1. Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, 60612, IL, USA

    Wilma A. Hofmann, Ljuba Stojiljkovic, Beata Fuchsova, Gabriela M. Vargas, Evangelos Mavrommatis & Primal de Lanerolle

  2. Department of Cell Ultrastructure and Molecular Biology, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague, 142 20, Czech Republic

    Vlada Philimonenko, Katarina Kysela & Pavel Hozak

  3. Department of Chemistry and Biochemistry, University of Colorado, Boulder, 80309, CO, USA

    James A. Goodrich

  4. Division of Developmental Biology, Children's Hospital Research Foundation, Cincinnati, 45229, OH, USA

    James L. Lessard

  5. Department of Microbiology and Immunology, University of Illinois at Chicago, Chicago, 60612, IL, USA

    Thomas J. Hope

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Hofmann, W., Stojiljkovic, L., Fuchsova, B. et al. Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II. Nat Cell Biol 6, 1094–1101 (2004). https://doi.org/10.1038/ncb1182

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