Synthetic lethality of rpn11-1 rpn10Δ is linked to altered proteasome assembly and activity

Abhishek Chandra, Li Chen, Kiran Madura

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

An rpn11-1 temperature-sensitive mutant shows defect in proteolysis, mitochondrial function and proteasome assembly. The Rpn11 protein is a proteasome subunit that deubiquitinates proteolytic substrates. Multiubiquitinated proteins interact with proteasome receptors, such as Rpn10, which intriguingly is also required for promoting proteasome stability. We report here that Rpn10 binds Rpn11, and genetic studies revealed synthetic lethality of an rpn11-1 rpn10Δ double mutant. The carboxy-terminus of Rpn11 is critical for function, as deletion of 7 C-terminal residues prevented suppression of rpn11-1 rpn10Δ. Native gel electrophoresis showed increased levels of the proteasome 20S catalytic particle in rpn11-1 rpn10Δ, and altered assembly. The inviability of rpn11-1 rpn10Δ was suppressed by rpn10uim, a mutant that can bind the proteasome, but not multiubiquitin chains. rpn10uim reduced the levels of free 20S, and increased formation of intact proteasomes. In contrast, rpn10vwa, which binds multiubiquitin chains but not the proteasome, failed to suppress rpn11-1 rpn10Δ. Moreover, high levels of multiubiquitinated proteins were bound to rpn10vwa, but were not delivered to the proteasome. Based on these findings, we propose that the lethality of rpn11-1 rpn10Δ results primarily from altered proteasome integrity. It is conceivable that Rpn10/Rpn11 interaction couples proteasome assembly to substrate binding.

Original languageEnglish (US)
Pages (from-to)543-557
Number of pages15
JournalCurrent Genetics
Volume56
Issue number6
DOIs
StatePublished - Dec 1 2010
Externally publishedYes

Fingerprint

Proteasome Endopeptidase Complex
Synthetic Lethal Mutations
Proteins
Proteolysis
Electrophoresis
Gels
Temperature

Keywords

  • Proteasome
  • Proteolysis
  • Rpn10
  • Rpn11
  • Ubiquitin

ASJC Scopus subject areas

  • Genetics

Cite this

Synthetic lethality of rpn11-1 rpn10Δ is linked to altered proteasome assembly and activity. / Chandra, Abhishek; Chen, Li; Madura, Kiran.

In: Current Genetics, Vol. 56, No. 6, 01.12.2010, p. 543-557.

Research output: Contribution to journalArticle

@article{a84187a2f0334446b2ccfd610b9924b2,
title = "Synthetic lethality of rpn11-1 rpn10Δ is linked to altered proteasome assembly and activity",
abstract = "An rpn11-1 temperature-sensitive mutant shows defect in proteolysis, mitochondrial function and proteasome assembly. The Rpn11 protein is a proteasome subunit that deubiquitinates proteolytic substrates. Multiubiquitinated proteins interact with proteasome receptors, such as Rpn10, which intriguingly is also required for promoting proteasome stability. We report here that Rpn10 binds Rpn11, and genetic studies revealed synthetic lethality of an rpn11-1 rpn10Δ double mutant. The carboxy-terminus of Rpn11 is critical for function, as deletion of 7 C-terminal residues prevented suppression of rpn11-1 rpn10Δ. Native gel electrophoresis showed increased levels of the proteasome 20S catalytic particle in rpn11-1 rpn10Δ, and altered assembly. The inviability of rpn11-1 rpn10Δ was suppressed by rpn10uim, a mutant that can bind the proteasome, but not multiubiquitin chains. rpn10uim reduced the levels of free 20S, and increased formation of intact proteasomes. In contrast, rpn10vwa, which binds multiubiquitin chains but not the proteasome, failed to suppress rpn11-1 rpn10Δ. Moreover, high levels of multiubiquitinated proteins were bound to rpn10vwa, but were not delivered to the proteasome. Based on these findings, we propose that the lethality of rpn11-1 rpn10Δ results primarily from altered proteasome integrity. It is conceivable that Rpn10/Rpn11 interaction couples proteasome assembly to substrate binding.",
keywords = "Proteasome, Proteolysis, Rpn10, Rpn11, Ubiquitin",
author = "Abhishek Chandra and Li Chen and Kiran Madura",
year = "2010",
month = "12",
day = "1",
doi = "10.1007/s00294-010-0321-3",
language = "English (US)",
volume = "56",
pages = "543--557",
journal = "Current Genetics",
issn = "0172-8083",
publisher = "Springer Verlag",
number = "6",

}

TY - JOUR

T1 - Synthetic lethality of rpn11-1 rpn10Δ is linked to altered proteasome assembly and activity

AU - Chandra, Abhishek

AU - Chen, Li

AU - Madura, Kiran

PY - 2010/12/1

Y1 - 2010/12/1

N2 - An rpn11-1 temperature-sensitive mutant shows defect in proteolysis, mitochondrial function and proteasome assembly. The Rpn11 protein is a proteasome subunit that deubiquitinates proteolytic substrates. Multiubiquitinated proteins interact with proteasome receptors, such as Rpn10, which intriguingly is also required for promoting proteasome stability. We report here that Rpn10 binds Rpn11, and genetic studies revealed synthetic lethality of an rpn11-1 rpn10Δ double mutant. The carboxy-terminus of Rpn11 is critical for function, as deletion of 7 C-terminal residues prevented suppression of rpn11-1 rpn10Δ. Native gel electrophoresis showed increased levels of the proteasome 20S catalytic particle in rpn11-1 rpn10Δ, and altered assembly. The inviability of rpn11-1 rpn10Δ was suppressed by rpn10uim, a mutant that can bind the proteasome, but not multiubiquitin chains. rpn10uim reduced the levels of free 20S, and increased formation of intact proteasomes. In contrast, rpn10vwa, which binds multiubiquitin chains but not the proteasome, failed to suppress rpn11-1 rpn10Δ. Moreover, high levels of multiubiquitinated proteins were bound to rpn10vwa, but were not delivered to the proteasome. Based on these findings, we propose that the lethality of rpn11-1 rpn10Δ results primarily from altered proteasome integrity. It is conceivable that Rpn10/Rpn11 interaction couples proteasome assembly to substrate binding.

AB - An rpn11-1 temperature-sensitive mutant shows defect in proteolysis, mitochondrial function and proteasome assembly. The Rpn11 protein is a proteasome subunit that deubiquitinates proteolytic substrates. Multiubiquitinated proteins interact with proteasome receptors, such as Rpn10, which intriguingly is also required for promoting proteasome stability. We report here that Rpn10 binds Rpn11, and genetic studies revealed synthetic lethality of an rpn11-1 rpn10Δ double mutant. The carboxy-terminus of Rpn11 is critical for function, as deletion of 7 C-terminal residues prevented suppression of rpn11-1 rpn10Δ. Native gel electrophoresis showed increased levels of the proteasome 20S catalytic particle in rpn11-1 rpn10Δ, and altered assembly. The inviability of rpn11-1 rpn10Δ was suppressed by rpn10uim, a mutant that can bind the proteasome, but not multiubiquitin chains. rpn10uim reduced the levels of free 20S, and increased formation of intact proteasomes. In contrast, rpn10vwa, which binds multiubiquitin chains but not the proteasome, failed to suppress rpn11-1 rpn10Δ. Moreover, high levels of multiubiquitinated proteins were bound to rpn10vwa, but were not delivered to the proteasome. Based on these findings, we propose that the lethality of rpn11-1 rpn10Δ results primarily from altered proteasome integrity. It is conceivable that Rpn10/Rpn11 interaction couples proteasome assembly to substrate binding.

KW - Proteasome

KW - Proteolysis

KW - Rpn10

KW - Rpn11

KW - Ubiquitin

UR - http://www.scopus.com/inward/record.url?scp=78649328672&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649328672&partnerID=8YFLogxK

U2 - 10.1007/s00294-010-0321-3

DO - 10.1007/s00294-010-0321-3

M3 - Article

C2 - 20941496

AN - SCOPUS:78649328672

VL - 56

SP - 543

EP - 557

JO - Current Genetics

JF - Current Genetics

SN - 0172-8083

IS - 6

ER -