The Structural Role of RPN10 in the 26S Proteasome and an RPN2-Binding Residue on RPN13 Are Functionally Important in Arabidopsis.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI, [2000-

Proteasome Endopeptidase Complex*/metabolism ; Arabidopsis*/genetics ; Arabidopsis*/metabolism ; Arabidopsis Proteins*/metabolism ; Arabidopsis Proteins*/genetics ; Arabidopsis Proteins*/chemistry ; Protein Binding*; Binding Sites ; Ubiquitin/metabolism ; Endopeptidases

The ubiquitin receptors RPN10 and RPN13 harbor multiple activities including ubiquitin binding; however, solid evidence connecting a particular activity to specific in vivo functions is scarce. Through complementation, the ubiquitin-binding site-truncated Arabidopsis RPN10 (N215) rescued the growth defects of rpn10-2 , supporting the idea that the ubiquitin-binding ability of RPN10 is dispensable and N215, which harbors a vWA domain, is fully functional. Instead, a structural role played by RPN10 in the 26S proteasomes is likely vital in vivo. A site-specific variant, RPN10-11A, that likely has a destabilized vWA domain could partially rescue the rpn10-2 growth defects and is not integrated into 26S proteasomes. Native polyacrylamide gel electrophoresis and mass spectrometry with rpn10-2 26S proteasomes showed that the loss of RPN10 reduced the abundance of double-capped proteasomes, induced the integration of specific subunit paralogues, and increased the association of ECM29, a well-known factor critical for quality checkpoints by binding and inhibiting aberrant proteasomes. Extensive Y2H and GST-pulldown analyses identified RPN2-binding residues on RPN13 that overlapped with ubiquitin-binding and UCH2-binding sites in the RPN13 C-terminus (246-254). Interestingly, an analysis of homozygous rpn10-2 segregation in a rpn13-1 background harboring RPN13 variants defective for ubiquitin binding and/or RPN2 binding supports the criticality of the RPN13-RPN2 association in vivo.

J Biol Chem. 2011 Oct 21;286(42):36641-51. (PMID: 21878651)
Mol Cell. 2009 Aug 14;35(3):280-90. (PMID: 19683493)
Nature. 2008 May 22;453(7194):548-52. (PMID: 18497827)
Plant Cell. 2003 Apr;15(4):965-80. (PMID: 12671091)
J Cell Sci. 2003 Mar 15;116(Pt 6):1023-33. (PMID: 12584246)
EMBO J. 2006 Dec 13;25(24):5742-53. (PMID: 17139257)
EMBO J. 2001 Dec 17;20(24):7096-107. (PMID: 11742986)
Mol Cell. 2002 Sep;10(3):495-507. (PMID: 12408819)
Trends Plant Sci. 2010 Jul;15(7):375-86. (PMID: 20399133)
J Cell Biol. 2000 Jul 10;150(1):119-30. (PMID: 10893261)
Plant Cell. 2011 Jul;23(7):2754-73. (PMID: 21764993)
Mol Cell. 2015 Mar 5;57(5):901-911. (PMID: 25702872)
J Biol Chem. 2017 Sep 29;292(39):16310-16320. (PMID: 28821611)
Sci Rep. 2019 Nov 19;9(1):17030. (PMID: 31745110)
Plant J. 2007 Aug;51(3):441-57. (PMID: 17559514)
Mol Cell. 2010 Jun 25;38(6):879-88. (PMID: 20620957)
Proc Natl Acad Sci U S A. 2012 Jan 31;109(5):1479-84. (PMID: 22215586)
J Biol Chem. 2004 Feb 20;279(8):6401-13. (PMID: 14623884)
Cell Rep. 2023 Jul 25;42(7):112701. (PMID: 37384533)
Plant Cell. 2016 Jun;28(6):1279-96. (PMID: 27194708)
Cell. 1998 Sep 4;94(5):615-23. (PMID: 9741626)
Genetics. 1998 Jun;149(2):677-92. (PMID: 9611183)
Plant J. 1998 Dec;16(6):735-43. (PMID: 10069079)
EMBO J. 2006 Oct 4;25(19):4524-36. (PMID: 16990800)
EMBO J. 2007 May 2;26(9):2339-49. (PMID: 17431397)
J Biol Chem. 2010 Aug 13;285(33):25554-69. (PMID: 20516081)
Bioinformatics. 2014 Aug 1;30(15):2114-20. (PMID: 24695404)
Nature. 2012 Jan 11;482(7384):186-91. (PMID: 22237024)
J Biol Chem. 2004 Dec 24;279(52):54849-61. (PMID: 15496406)
Annu Rev Biochem. 2012;81:203-29. (PMID: 22524316)
Nature. 1997 Feb 20;385(6618):737-40. (PMID: 9034192)
Proc Natl Acad Sci U S A. 2009 Jul 21;106(29):11943-7. (PMID: 19581588)
J Biol Chem. 2013 Oct 11;288(41):29467-81. (PMID: 23995839)
Mol Cell. 2015 Mar 5;57(5):887-900. (PMID: 25702870)
Science. 2016 Feb 19;351(6275):. (PMID: 26912900)
EMBO Rep. 2008 Dec;9(12):1237-43. (PMID: 18927584)
Nat Cell Biol. 2006 Sep;8(9):994-1002. (PMID: 16906146)
Plant Cell. 1990 Aug;2(8):755-67. (PMID: 2152125)
Genome Biol. 2010;11(3):R25. (PMID: 20196867)
Mol Cell. 2015 Jun 18;58(6):1053-66. (PMID: 26004230)
J Biol Chem. 1998 Apr 24;273(17):10396-401. (PMID: 9553097)
Front Mol Biosci. 2019 Jun 07;6:40. (PMID: 31231659)
Mol Biol Cell. 2000 Oct;11(10):3425-39. (PMID: 11029046)
J Biol Chem. 2019 Nov 15;294(46):17570-17592. (PMID: 31562246)
Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):20992-7. (PMID: 21098295)
J Biol Chem. 1994 Mar 11;269(10):7059-61. (PMID: 8125911)
J Biol Chem. 2011 Oct 21;286(42):36652-66. (PMID: 21878652)
Nat Methods. 2012 Mar 04;9(4):357-9. (PMID: 22388286)
Nature. 2008 May 22;453(7194):481-8. (PMID: 18497817)
Plant J. 2017 Feb;89(4):789-804. (PMID: 27862469)
J Biol Chem. 2007 Nov 30;282(48):34869-76. (PMID: 17911101)
Proc Natl Acad Sci U S A. 2012 Jan 3;109(1):149-54. (PMID: 22187461)
Genome Res. 2002 Apr;12(4):656-64. (PMID: 11932250)
Anal Chem. 1996 Mar 1;68(5):850-8. (PMID: 8779443)
Mol Cell. 2010 May 14;38(3):404-15. (PMID: 20471946)
Methods Mol Biol. 2014;1156:213-22. (PMID: 24791991)
Mol Cell. 2010 Nov 24;40(4):671-81. (PMID: 21095592)
Dev Growth Differ. 2001 Feb;43(1):25-31. (PMID: 11148449)
Mol Cell Biol. 2007 Oct;27(19):6629-38. (PMID: 17646385)
Mol Cell Biol. 1996 Nov;16(11):6020-8. (PMID: 8887631)
FEBS J. 2010 Feb;277(3):796-816. (PMID: 20059542)
Nat Rev Mol Cell Biol. 2009 Jun;10(6):385-97. (PMID: 19424292)
J Biol Chem. 2016 Jun 17;291(25):13147-59. (PMID: 27129254)
J Biol Chem. 1998 Jan 23;273(4):1970-81. (PMID: 9442033)
Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):856-60. (PMID: 8570648)
PLoS Genet. 2015 Jul 29;11(7):e1005401. (PMID: 26222436)
Plant Signal Behav. 2012 Jul;7(7):722-7. (PMID: 22751321)
EMBO J. 2013 Oct 16;32(20):2697-707. (PMID: 23982732)

Keywords: 26S proteasome; Arabidopsis; ECM29; PA200; RPN10; RPN13; RPN2; UCH1; UCH2; ubiquitin receptor

EC 3.4.25.1 (Proteasome Endopeptidase Complex)
0 (Arabidopsis Proteins)
EC 3.4.99.- (ATP dependent 26S protease)
0 (RPN10 protein, Arabidopsis)
EC 3.4.99.- (UBP13 protein, Arabidopsis)
0 (Ubiquitin)
EC 3.4.- (Endopeptidases)

Date Created: 20241109 Date Completed: 20241109 Latest Revision: 20241116

20241116

PMC11546751

10.3390/ijms252111650

39519207