title: A photoconvertible reporter of the ubiquitin-proteasome system in vivo.
abstract: The ubiquitin-proteasome system (UPS) orchestrates many cellular and tissue-specific processes by degrading damaged and key regulatory proteins. To enable investigation of UPS activity in different cell types in a living animal, we developed a photoconvertible fluorescent UPS reporter system for live imaging and quantification of protein degradation in Caenorhabditis elegans. Our reporter consists of the photoconvertible fluorescent protein Dendra2 targeted for proteasomal degradation by fusion to the UbG76V mutant form of ubiquitin. In contrast to previous reporters, this system permits quantification of UPS activity independently of protein synthesis. Our reporter revealed that UPS-mediated protein degradation varies in a cell type-specific and age-dependent manner in C. elegans.
authors.affiliation: Molecular Cancer Biology Program and Institute for Biomedicine, Biomedicum Helsinki, University of Helsinki, Helsinki, Finland.
authors.forename: Geert
authors.initials: G
authors.lastname: Hamer
authors.forename: Olli
authors.initials: O
authors.lastname: Matilainen
authors.forename: Carina I
authors.initials: CI
authors.lastname: Holmberg
mesh_terms: D000375:Aging / Q000378:metabolism; D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000502:physiology; D004298:Dopamine / Q000502:physiology; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D046988:Proteasome Endopeptidase Complex / Q000502:physiology*; D034622:RNA Interference; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology; D025801:Ubiquitin / Q000378:metabolism*; D005680:gamma-Aminobutyric Acid / Q000502:physiology
chemical_list: D029742:Caenorhabditis elegans Proteins; D008164:Luminescent Proteins; C516341:RPN-10 protein, C elegans; D018160:Receptors, Cytoplasmic and Nuclear; D025801:Ubiquitin; D005680:gamma-Aminobutyric Acid; D046988:Proteasome Endopeptidase Complex; D004298:Dopamine
references.citation: Mol Biol Cell. 2006 Dec;17(12):5356-71
references.pmid: 17050737
references.citation: FASEB J. 2009 Jan;23(1):123-33
references.pmid: 18796559
references.citation: Science. 2001 May 25;292(5521):1552-5
references.pmid: 11375494
references.citation: Mol Biol Cell. 2004 Feb;15(2):657-64
references.pmid: 14668486
references.citation: Biol Chem. 2008 Mar;389(3):203-9
references.pmid: 18208355
references.citation: EMBO J. 2004 Oct 27;23(21):4307-18
references.pmid: 15470501
references.citation: Genetics. 1974 May;77(1):71-94
references.pmid: 4366476
references.citation: J Biol Chem. 2009 Oct 9;284(41):28253-28262
references.pmid: 19679666
references.citation: Biotechniques. 2007 Apr;42(4):446, 448, 450
references.pmid: 17489230
references.citation: EMBO J. 2009 Jul 8;28(13):1867-77
references.pmid: 19387488
references.citation: EMBO J. 1992 Feb;11(2):497-505
references.pmid: 1311250
references.citation: Proc Natl Acad Sci U S A. 2008 May 13;105(19):7094-9
references.pmid: 18467495
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Dev Cell. 2007 Feb;12(2):235-46
references.pmid: 17276341
references.citation: FASEB J. 2005 Dec;19(14):2051-3
references.pmid: 16188962
references.citation: FEBS Lett. 2007 Jun 19;581(15):2854-61
references.pmid: 17418826
references.citation: Aging Cell. 2007 Oct;6(5):599-606
references.pmid: 17681036
references.citation: FEBS Lett. 2002 Apr 10;516(1-3):53-7
references.pmid: 11959102
references.citation: Science. 2009 May 29;324(5931):1196-8
references.pmid: 19372390
references.citation: Proc Natl Acad Sci U S A. 2007 Apr 3;104(14):5947-52
references.pmid: 17392428
references.citation: Nat Biotechnol. 2003 Aug;21(8):897-902
references.pmid: 12872133
references.citation: Nat Protoc. 2007;2(8):2024-32
references.pmid: 17703215
references.citation: Nat Biotechnol. 2000 May;18(5):538-43
references.pmid: 10802622
references.citation: J Neurosci. 2006 Jul 19;26(29):7597-606
references.pmid: 16855087
references.citation: PLoS One. 2009 Jun 11;4(6):e5888
references.pmid: 19517018
references.citation: EMBO J. 1991 Dec;10(12):3959-70
references.pmid: 1935914
references.citation: Aging Cell. 2003 Apr;2(2):131-9
references.pmid: 12882326
references.citation: Gene. 2001 Jan 24;263(1-2):103-12
references.pmid: 11223248
references.citation: Bioessays. 2008 Nov;30(11-12):1172-84
references.pmid: 18937370
references.citation: Nat Biotechnol. 2000 Dec;18(12):1298-302
references.pmid: 11101811
references.citation: J Biol Chem. 1994 Oct 28;269(43):27021-8
references.pmid: 7929443
references.citation: Ann Neurol. 2006 Aug;60(2):260-4
references.pmid: 16862585
references.citation: Mol Cell Biol. 1996 Nov;16(11):6020-8
references.pmid: 8887631
references.citation: Science. 2003 May 16;300(5622):1142-5
references.pmid: 12750521
references.citation: Nature. 2002 Oct 24;419(6909):808-14
references.pmid: 12397350