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Isolation and Identification of PACE-Binding Protein Rpn4, a New Transcriptional Activator Regulating 26S-Proteasomal and Other Genes

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Abstract

A new upstream activating sequence (UAS) 5"-GGTGGCAAA-3" was detected in the promoters of 27 out of the 33 proteasomal genes and of several genes related to the ubiquitin-proteasomal system of Saccharomyces cerevisiae. The sequence was termed proteasome-associated control element (PACE). Gel retardation assay revealed specific binding of PACE with an extracted protein. The protein (64K) was purified by affinity chromatography and was homogeneous by SDS-PAGE. Microsequencing showed that the protein is Rpn4p. The ability of Rpn4p to activate transcription was demonstrated with constructs containing fragments of the RPN5and CDC48gene promoters and reporter cat. Binding to PACE, Rpn4p may act as a common transcription factor on the proteasomal and proteasome-related genes.

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Authors and Affiliations

  1. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 117984, Russia

    A. B. Kapranov, M. V. Kuryatova, O. V. Preobrazhenskaya, V. V. Tutyaeva & V. L. Karpov

  2. Adolf-Butenandt-Institut der, Ludwig-Maximilians-Univrsität, München, D-80336, Germany

    R. Stucka & H. Feldmann

Authors
  1. A. B. Kapranov
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  2. M. V. Kuryatova
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  3. O. V. Preobrazhenskaya
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  4. V. V. Tutyaeva
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  5. R. Stucka
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  6. H. Feldmann
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  7. V. L. Karpov
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Kapranov, A.B., Kuryatova, M.V., Preobrazhenskaya, O.V. et al. Isolation and Identification of PACE-Binding Protein Rpn4, a New Transcriptional Activator Regulating 26S-Proteasomal and Other Genes. Molecular Biology 35, 356–364 (2001). https://doi.org/10.1023/A:1010470627747

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  • DOI: https://doi.org/10.1023/A:1010470627747

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