The lysine 48 and lysine 63 ubiquitin conjugates are processed differently by the 26 s proteasome
The role of Lys-63 ubiquitin chains in targeting proteins for proteasomal degradation is still obscure. We systematically compared proteasomal processing of Lys-63 ubiquitin chains with that of the canonical proteolytic signal, Lys-48 ubiquitin chains. Quantitative mass spectrometric analysis of ubiquitin chains in HeLa cells determines that the levels of Lys-63 ubiquitin chains are insensitive to short-time proteasome inhibition. Also, the Lys-48/Lys-63 ratio in the 26 S proteasome-bound fraction is 1.7-fold more than that in the cell lysates, likely because some cellular Lys-63 ubiquitin conjugates are sequestered by Lys-63 chain-specific binding proteins. In vitro, Lys-48 and Lys-63 ubiquitin chains bind the 26 S proteasome comparably, whereas Lys-63 chains are deubiquitinated 6-fold faster than Lys-48 chains. Also, Lys-63 tetraubiquitin-conjugated UbcH10 is rapidly deubiquitinated into the monoubiquitinated form, whereas Lys-48 tetraubiquitin targets UbcH10 for degradation. Furthermore, we found that both the ubiquitin aldehyde- and 1,10-phenanthroline-sensitive deubiquitinating activities of the 26 S proteasome contribute to Lys-48- and Lys-63-linkage deubiquitination, albeit the inhibitory extents are different. Together, our findings suggest that compared with Lys-48 chains, cellular Lys-63 chains have less proteasomal accessibility, and proteasome-bound Lys-63 chains are more rapidly deubiquitinated, which could cause inefficient degradation of Lys-63 conjugates.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2009 |
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| Erschienen: |
2009 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:284 |
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| Enthalten in: |
The Journal of biological chemistry - 284(2009), 51 vom: 18. Dez., Seite 35485-94 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Jacobson, Andrew D [VerfasserIn] |
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| Links: |
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| Themen: |
ATP dependent 26S protease |
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| Anmerkungen: |
Date Completed 06.01.2010 Date Revised 16.03.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.1074/jbc.M109.052928 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM192314092 |
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| 100 | 1 | |a Jacobson, Andrew D |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The lysine 48 and lysine 63 ubiquitin conjugates are processed differently by the 26 s proteasome |
| 264 | 1 | |c 2009 | |
| 336 | |a Text |b txt |2 rdacontent | ||
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| 500 | |a Date Completed 06.01.2010 | ||
| 500 | |a Date Revised 16.03.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a The role of Lys-63 ubiquitin chains in targeting proteins for proteasomal degradation is still obscure. We systematically compared proteasomal processing of Lys-63 ubiquitin chains with that of the canonical proteolytic signal, Lys-48 ubiquitin chains. Quantitative mass spectrometric analysis of ubiquitin chains in HeLa cells determines that the levels of Lys-63 ubiquitin chains are insensitive to short-time proteasome inhibition. Also, the Lys-48/Lys-63 ratio in the 26 S proteasome-bound fraction is 1.7-fold more than that in the cell lysates, likely because some cellular Lys-63 ubiquitin conjugates are sequestered by Lys-63 chain-specific binding proteins. In vitro, Lys-48 and Lys-63 ubiquitin chains bind the 26 S proteasome comparably, whereas Lys-63 chains are deubiquitinated 6-fold faster than Lys-48 chains. Also, Lys-63 tetraubiquitin-conjugated UbcH10 is rapidly deubiquitinated into the monoubiquitinated form, whereas Lys-48 tetraubiquitin targets UbcH10 for degradation. Furthermore, we found that both the ubiquitin aldehyde- and 1,10-phenanthroline-sensitive deubiquitinating activities of the 26 S proteasome contribute to Lys-48- and Lys-63-linkage deubiquitination, albeit the inhibitory extents are different. Together, our findings suggest that compared with Lys-48 chains, cellular Lys-63 chains have less proteasomal accessibility, and proteasome-bound Lys-63 chains are more rapidly deubiquitinated, which could cause inefficient degradation of Lys-63 conjugates | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Ubiquitin |2 NLM | |
| 650 | 7 | |a Proteasome Endopeptidase Complex |2 NLM | |
| 650 | 7 | |a EC 3.4.25.1 |2 NLM | |
| 650 | 7 | |a ATP dependent 26S protease |2 NLM | |
| 650 | 7 | |a EC 3.4.99.- |2 NLM | |
| 650 | 7 | |a Lysine |2 NLM | |
| 650 | 7 | |a K3Z4F929H6 |2 NLM | |
| 700 | 1 | |a Zhang, Nan-Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Xu, Ping |e verfasserin |4 aut | |
| 700 | 1 | |a Han, Ke-Jun |e verfasserin |4 aut | |
| 700 | 1 | |a Noone, Seth |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Junmin |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Chang-Wei |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t The Journal of biological chemistry |d 1945 |g 284(2009), 51 vom: 18. Dez., Seite 35485-94 |w (DE-627)NLM000004995 |x 1083-351X |7 nnns |
| 773 | 1 | 8 | |g volume:284 |g year:2009 |g number:51 |g day:18 |g month:12 |g pages:35485-94 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1074/jbc.M109.052928 |3 Volltext |
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