Proteasomal ATPase-associated factor 1 negatively regulates proteasome activity by interacting with proteasomal ATPases

Yoon Park, Yong Pil Hwang, Jong Sik Lee, Sang Hyun Seo, Kim Yoon Sungjoo, Jong-Bok Yoon

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

The 26S proteasome, composed of the 20S core and the 19S regulatory complex, plays a central role in ubiquitin-dependent proteolysis by catalyzing degradation of polyubiquitinated proteins. In a search for proteins involved in regulation of the proteasome, we affinity purified the 19S regulatory complex from HeLa cells and identified a novel protein of 43 kDa in size as an associated protein. Immunoprecipitation analyses suggested that this protein specifically interacted with the proteasomal ATPases. Hence the protein was named proteasomal ATPase-associated factor 1 (PAAF1). Immunoafinity purification of PAAF1 confirmed its interaction with the 19S regulatory complex and further showed that the 19S regulatory complex bound with PAAF1 was not stably associated with the 20S core. Overexpression of PAAF1 in HeLa cells decreased the level of the 20S core associated with the 19S complex in a dose-dependent fashion, suggesting that PAAF1 binding to proteasomal ATPases inhibited the assembly of the 26S proteasome. Proteasomal degradation assays using reporters based on green fluorescent protein revealed that overexpression of PAAF1 inhibited the proteasome activity in vivo. Furthermore, the suppression of PAAF1 expression that is mediated by small inhibitory RNA enhanced the proteasome activity. These results suggest that PAAF1 functions as a negative regulator of the proteasome by controlling the assembly/disassembly of the proteasome.

Original languageEnglish
Pages (from-to)3842-3853
Number of pages12
JournalMolecular and Cellular Biology
Volume25
Issue number9
DOIs
Publication statusPublished - 2005 May 1

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Proteasome Endopeptidase Complex
Adenosine Triphosphatases
Proteins
HeLa Cells
Proteolysis
Ubiquitin
Green Fluorescent Proteins
Immunoprecipitation
RNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Park, Yoon ; Hwang, Yong Pil ; Lee, Jong Sik ; Seo, Sang Hyun ; Sungjoo, Kim Yoon ; Yoon, Jong-Bok. / Proteasomal ATPase-associated factor 1 negatively regulates proteasome activity by interacting with proteasomal ATPases. In: Molecular and Cellular Biology. 2005 ; Vol. 25, No. 9. pp. 3842-3853.
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Proteasomal ATPase-associated factor 1 negatively regulates proteasome activity by interacting with proteasomal ATPases. / Park, Yoon; Hwang, Yong Pil; Lee, Jong Sik; Seo, Sang Hyun; Sungjoo, Kim Yoon; Yoon, Jong-Bok.

In: Molecular and Cellular Biology, Vol. 25, No. 9, 01.05.2005, p. 3842-3853.

Research output: Contribution to journalArticle

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