TY - JOUR
T1 - The 19S regulatory particle of the proteasome is required for efficient transcription elongation by RNA polymerase II
AU - Ferdous, Anwarul
AU - Gonzalez, Fernando
AU - Sun, Liping
AU - Kodadek, Thomas
AU - Johnston, Stephen Albert
N1 - Funding Information:
We thank Steven J. Russell for constructing the sug1-20 and sug1-25 strains and for producing antibodies against Sug1 and Sug2. Sincere thanks to Dieter Wolf, Ray J. Deshaies, and Gerald C. Johnston for strains, and Keiji Tanaka (anti-20S), Tim Formosa (anti-Cdc68), and Jonathan C. Swaffield (anti-Rtp5) for antibodies. We also thank Prof. Deshaies for providing us with a protocol, prior to publication, for immunoaffinity purification of epitope-tagged proteasome. Modification of this protocol resulted in the method employed to prepare the 26S and 19S complexes used in this study. We also thank Mark Combs for mass spectrometric analysis of the 19S and 26S proteasome, Xiang Chen, Eunice Webb, and Zakir Siddiquee for their technical expertise, and members of S.A.J.'s and T.K.'s laboratories for helpful comments on the manuscript. This work was supported by a grant from the American Cancer Society to T.K. and unrestricted funds to S.A.J.
PY - 2001/5/25
Y1 - 2001/5/25
N2 - It is generally thought that the primary or even sole activity of the 19S regulatory particle of the 26S proteasome is to facilitate the degradation of polyubiquitinated proteins by the 20S-core subunit. However, we present evidence that the 19S complex is required for efficient elongation of RNA polymerase II (RNAP II) in vitro and in vivo. First, yeast strains carrying alleles of SUG1 and SUG2, encoding 19S components, exhibit phenotypes indicative of elongation defects. Second, in vitro transcription is inhibited by antibodies raised against Sug1, or by heat-inactivating temperature-sensitive Sug1 mutants with restoration of elongation by addition of immunopurified 19S complex. Finally, Cdc68, a known elongation factor, coimmunoprecipitates with the 19S complex, indicating a physical interaction. Inhibition of the 20S proteolytic core of the proteasome has no effect on elongation. This work defines a nonproteolytic role for the 19S complex in RNAP II transcription.
AB - It is generally thought that the primary or even sole activity of the 19S regulatory particle of the 26S proteasome is to facilitate the degradation of polyubiquitinated proteins by the 20S-core subunit. However, we present evidence that the 19S complex is required for efficient elongation of RNA polymerase II (RNAP II) in vitro and in vivo. First, yeast strains carrying alleles of SUG1 and SUG2, encoding 19S components, exhibit phenotypes indicative of elongation defects. Second, in vitro transcription is inhibited by antibodies raised against Sug1, or by heat-inactivating temperature-sensitive Sug1 mutants with restoration of elongation by addition of immunopurified 19S complex. Finally, Cdc68, a known elongation factor, coimmunoprecipitates with the 19S complex, indicating a physical interaction. Inhibition of the 20S proteolytic core of the proteasome has no effect on elongation. This work defines a nonproteolytic role for the 19S complex in RNAP II transcription.
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U2 - 10.1016/S1097-2765(01)00250-7
DO - 10.1016/S1097-2765(01)00250-7
M3 - Article
C2 - 11389845
AN - SCOPUS:0035947238
SN - 1097-2765
VL - 7
SP - 981
EP - 991
JO - Molecular cell
JF - Molecular cell
IS - 5
ER -