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

Wilma A. Hofmann; Ljuba Stojiljkovic; Beata Fuchsova; Gabriela M. Vargas; Evangelos Mavrommatis; Vlada Philimonenko; Katarina Kysela; James A. Goodrich; James L. Lessard; Thomas J. Hope; Pavel Hozak; Primal de Lanerolle

doi:10.1038/ncb1182

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

Wilma A. Hofmann, Ljuba Stojiljkovic, Beata Fuchsova, Gabriela M. Vargas, Evangelos Mavrommatis, Vlada Philimonenko, Katarina Kysela, James A. Goodrich, James L. Lessard, Thomas J. Hope, Pavel Hozak, Primal de Lanerolle^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

320 Scopus citations

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.

Original language	English (US)
Pages (from-to)	1094-1101
Number of pages	8
Journal	Nature Cell Biology
Volume	6
Issue number	11
DOIs	https://doi.org/10.1038/ncb1182
State	Published - Nov 2004

Funding

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).

ASJC Scopus subject areas

Cell Biology

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@article{a1ad0eb4c671438bb34fde425fe8204c,

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

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.",

author = "Hofmann, {Wilma A.} and Ljuba Stojiljkovic and Beata Fuchsova and Vargas, {Gabriela M.} and Evangelos Mavrommatis and Vlada Philimonenko and Katarina Kysela and Goodrich, {James A.} and Lessard, {James L.} and Hope, {Thomas J.} and Pavel Hozak and {de Lanerolle}, Primal",

note = "Funding Information: 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).",

year = "2004",

month = nov,

doi = "10.1038/ncb1182",

language = "English (US)",

volume = "6",

pages = "1094--1101",

journal = "Nature Cell Biology",

issn = "1465-7392",

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T1 - Actin is part of pre-initiation complexes and is necessary for transcription by RNA polymerase II

AU - Hofmann, Wilma A.

AU - Stojiljkovic, Ljuba

AU - Fuchsova, Beata

AU - Vargas, Gabriela M.

AU - Mavrommatis, Evangelos

AU - Philimonenko, Vlada

AU - Kysela, Katarina

AU - Goodrich, James A.

AU - Lessard, James L.

AU - Hope, Thomas J.

AU - Hozak, Pavel

AU - de Lanerolle, Primal

N1 - Funding Information: 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).

PY - 2004/11

Y1 - 2004/11

N2 - 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.

AB - 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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JO - Nature Cell Biology

JF - Nature Cell Biology

IS - 11

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

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