Pyrrolidine dithiocarbamate and zinc inhibit proteasome-dependent proteolysis

Insook Kim, Chul Hoon Kim, Joo Hee Kim, Jinu Lee, Junjeong Choi, Zheng Ai Chen, Min Goo Lee, Kwang Chul Chung, Chung Y. Hsu, Young Soo Ahn

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

Proteasomes play important roles in a variety of cellular processes such as cell cycle progression, signal transduction and immune responses. Proteasome activity is important in maintaining rapid turnover of short-lived proteins, as well as preventing accumulation of misfolded or damaged proteins. Alteration in ubiquitin-proteasome function may be detrimental to its crucial role in maintaining cellular homeostasis. Here, we have found that treatment of pyrrolidine dithiocarbamate (PDTC), a zinc ionophore, resulted in the accumulation of several proteasome substrates including p53 and p21 in HeLa cells. The PDTC effect was due to an extended half-life of these proteins through the mobilization of zinc. PDTC and/or zinc also increased fluorescence intensity of UbG76V-GFP fusion protein that is degraded rapidly by the ubiquitin-proteasome system. Treatment of cells with zinc induced formation of ubiquitinated inclusions in the centrosome, a histological marker of proteasome inhibition. Western blotting showed zinc-induced increase in laddering bands of polyubiquitin-conjugated proteins. In vitro study, zinc inhibited the ubiquitin-independent proteasomal degradations of p21 and α-synuclein. These results suggest that zinc may modulate cell functions through its action on the turnover of proteins that are susceptible to proteasome-dependent proteolysis.

Original languageEnglish
Pages (from-to)229-238
Number of pages10
JournalExperimental Cell Research
Volume298
Issue number1
DOIs
Publication statusPublished - 2004 Aug 1

Fingerprint

Proteasome Endopeptidase Complex
Proteolysis
Zinc
Ubiquitin
Proteins
Synucleins
Polyubiquitin
Centrosome
Ionophores
pyrrolidine dithiocarbamic acid
HeLa Cells
Half-Life
Signal Transduction
Cell Cycle
Homeostasis
Fluorescence
Western Blotting

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Kim, Insook ; Kim, Chul Hoon ; Kim, Joo Hee ; Lee, Jinu ; Choi, Junjeong ; Chen, Zheng Ai ; Lee, Min Goo ; Chung, Kwang Chul ; Hsu, Chung Y. ; Ahn, Young Soo. / Pyrrolidine dithiocarbamate and zinc inhibit proteasome-dependent proteolysis. In: Experimental Cell Research. 2004 ; Vol. 298, No. 1. pp. 229-238.
@article{f502379bb0f042c0ad7b68c1c3d68bf7,
title = "Pyrrolidine dithiocarbamate and zinc inhibit proteasome-dependent proteolysis",
abstract = "Proteasomes play important roles in a variety of cellular processes such as cell cycle progression, signal transduction and immune responses. Proteasome activity is important in maintaining rapid turnover of short-lived proteins, as well as preventing accumulation of misfolded or damaged proteins. Alteration in ubiquitin-proteasome function may be detrimental to its crucial role in maintaining cellular homeostasis. Here, we have found that treatment of pyrrolidine dithiocarbamate (PDTC), a zinc ionophore, resulted in the accumulation of several proteasome substrates including p53 and p21 in HeLa cells. The PDTC effect was due to an extended half-life of these proteins through the mobilization of zinc. PDTC and/or zinc also increased fluorescence intensity of UbG76V-GFP fusion protein that is degraded rapidly by the ubiquitin-proteasome system. Treatment of cells with zinc induced formation of ubiquitinated inclusions in the centrosome, a histological marker of proteasome inhibition. Western blotting showed zinc-induced increase in laddering bands of polyubiquitin-conjugated proteins. In vitro study, zinc inhibited the ubiquitin-independent proteasomal degradations of p21 and α-synuclein. These results suggest that zinc may modulate cell functions through its action on the turnover of proteins that are susceptible to proteasome-dependent proteolysis.",
author = "Insook Kim and Kim, {Chul Hoon} and Kim, {Joo Hee} and Jinu Lee and Junjeong Choi and Chen, {Zheng Ai} and Lee, {Min Goo} and Chung, {Kwang Chul} and Hsu, {Chung Y.} and Ahn, {Young Soo}",
year = "2004",
month = "8",
day = "1",
doi = "10.1016/j.yexcr.2004.04.017",
language = "English",
volume = "298",
pages = "229--238",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

Pyrrolidine dithiocarbamate and zinc inhibit proteasome-dependent proteolysis. / Kim, Insook; Kim, Chul Hoon; Kim, Joo Hee; Lee, Jinu; Choi, Junjeong; Chen, Zheng Ai; Lee, Min Goo; Chung, Kwang Chul; Hsu, Chung Y.; Ahn, Young Soo.

In: Experimental Cell Research, Vol. 298, No. 1, 01.08.2004, p. 229-238.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Pyrrolidine dithiocarbamate and zinc inhibit proteasome-dependent proteolysis

AU - Kim, Insook

AU - Kim, Chul Hoon

AU - Kim, Joo Hee

AU - Lee, Jinu

AU - Choi, Junjeong

AU - Chen, Zheng Ai

AU - Lee, Min Goo

AU - Chung, Kwang Chul

AU - Hsu, Chung Y.

AU - Ahn, Young Soo

PY - 2004/8/1

Y1 - 2004/8/1

N2 - Proteasomes play important roles in a variety of cellular processes such as cell cycle progression, signal transduction and immune responses. Proteasome activity is important in maintaining rapid turnover of short-lived proteins, as well as preventing accumulation of misfolded or damaged proteins. Alteration in ubiquitin-proteasome function may be detrimental to its crucial role in maintaining cellular homeostasis. Here, we have found that treatment of pyrrolidine dithiocarbamate (PDTC), a zinc ionophore, resulted in the accumulation of several proteasome substrates including p53 and p21 in HeLa cells. The PDTC effect was due to an extended half-life of these proteins through the mobilization of zinc. PDTC and/or zinc also increased fluorescence intensity of UbG76V-GFP fusion protein that is degraded rapidly by the ubiquitin-proteasome system. Treatment of cells with zinc induced formation of ubiquitinated inclusions in the centrosome, a histological marker of proteasome inhibition. Western blotting showed zinc-induced increase in laddering bands of polyubiquitin-conjugated proteins. In vitro study, zinc inhibited the ubiquitin-independent proteasomal degradations of p21 and α-synuclein. These results suggest that zinc may modulate cell functions through its action on the turnover of proteins that are susceptible to proteasome-dependent proteolysis.

AB - Proteasomes play important roles in a variety of cellular processes such as cell cycle progression, signal transduction and immune responses. Proteasome activity is important in maintaining rapid turnover of short-lived proteins, as well as preventing accumulation of misfolded or damaged proteins. Alteration in ubiquitin-proteasome function may be detrimental to its crucial role in maintaining cellular homeostasis. Here, we have found that treatment of pyrrolidine dithiocarbamate (PDTC), a zinc ionophore, resulted in the accumulation of several proteasome substrates including p53 and p21 in HeLa cells. The PDTC effect was due to an extended half-life of these proteins through the mobilization of zinc. PDTC and/or zinc also increased fluorescence intensity of UbG76V-GFP fusion protein that is degraded rapidly by the ubiquitin-proteasome system. Treatment of cells with zinc induced formation of ubiquitinated inclusions in the centrosome, a histological marker of proteasome inhibition. Western blotting showed zinc-induced increase in laddering bands of polyubiquitin-conjugated proteins. In vitro study, zinc inhibited the ubiquitin-independent proteasomal degradations of p21 and α-synuclein. These results suggest that zinc may modulate cell functions through its action on the turnover of proteins that are susceptible to proteasome-dependent proteolysis.

UR - http://www.scopus.com/inward/record.url?scp=3042745312&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3042745312&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2004.04.017

DO - 10.1016/j.yexcr.2004.04.017

M3 - Article

C2 - 15242777

AN - SCOPUS:3042745312

VL - 298

SP - 229

EP - 238

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -