REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
α-UBR5	Abcam	Cat#ab70311; RRID: AB_2210186
α-GFP (rabbit)	Sigma	Cat#G1544; RRID: AB_439690
α-GFP (mouse)	Sigma	Cat#G6539; RRID: AB_259941
α-Flag (mouse)	Sigma	Cat#F1804; RRID: AB_262044
α-Flag (rabbit)	Sigma	Cat#F7425; RRID: AB_439687
α-HA (rat)	Sigma	Cat#3F10
α-HA (rabbit)	Abcam	Cat#ab9110; RRID: AB_307019
α-Myc	Santa Cruz Biotechnology	Cat#sc-789; RRID: AB_631274
α-active β-catenin (ABC)	Cell Signaling Technologies	Cat#8814S; RRID: AB_11127203
α-β-catenin	BD Transduction Laboratories	Cat#610153
α-XIAP	BD Transduction Laboratories	Cat#610763; RRID: AB_398086
α-HUWE1	Abcam	Cat#ab70161; RRID: AB_1209511
α-TRIP12	Abcam	Cat#ab86220; RRID: AB_1925533
α-HECTD1	Abcam	Cat#ab101992; RRID: AB_10711075
α-UBE3C	Abcam	Cat#ab101512; RRID: AB_10711205
α-β-tubulin	Sigma	Cat#T4026; RRID: AB_477577
α-TLE1-4	Santa Cruz Biotechnology	Cat#sc-13373; RRID: AB_2203721
α-GST	Abcam	Cat#ab19256; RRID: AB_444809
HRP conjugated Goat α-Mouse	Santa Cruz Biotechnology	Cat#sc-2005; RRID: AB_631736
HRP conjugated Goat α-Rabbit	Santa Cruz Biotechnology	Cat#sc-2301; RRID: AB_650500
HRP conjugated Goat α-Rat	Santa Cruz Biotechnology	Cat#sc-2032; RRID: AB_631755
HRP conjugated Donkey?α-Goat	R&D Systems	Cat#HAF109; RRID: AB_357236
Alexa Fluor 488 conjugated Goat α-Rabbit	Life Technologies	Cat#A11008
Alexa Fluor 488 conjugated Goat α-Mouse	Life Technologies	Cat#A11029
Alexa Fluor 546 conjugated Goat α-Mouse	Life Technologies	Cat#A11003
Alexa Fluor 647 conjugated Goat α-Guinea pig	Invitrogen	Cat#A21450
α-Senseless	Prof. Hugo J. Bellen	N/A
α-Vestigial	Prof. Sean B. Carroll	N/A
α-Wingless	DSHB	Cat#4D4; RRID: AB_528512
α-Armadillo	DSHB	Cat#N27A1; RRID: AB_528089
Chemicals, Peptides, and Recombinant Proteins
Ni-NTA Agarose	QIAGEN	Cat#30210
α-FLAG M2 Affinity Gel	Sigma	Cat#A2220
EZview Red α-HA Affinity Gel	Sigma	Cat#E6779
GFP-trap_A	Chromotek	Cat#gta-20
Glutathione Sepharose 4b	GE Healthcare	Cat#17075601
Dynabeads MyOne Streptavidin C1	Invitrogen	Cat#65001
Lipofectamine2000	Invitrogen	Cat#11668019
Polyethylenimine, linear, MW25000	Polysciences	Cat#23966
EDTA-free Protease Inhibitor Cocktail	Roche	Cat#04693159001
MG132	Sigma	Cat#C2211
NMS-873	Cayman Chemical co.	Cat#1418013-75-8
CB-5083	Cayman Chemical co.	Cat#1542705-92-9
Cycloheximide	Sigma	Cat#C104450
Puromycin dihydrochloride	Sigma	Cat#P8833
3xFLAG-Peptide	Sigma	Cat#F4799
L-Glutathione reduced	Sigma	Cat#G4251
VectaShield with DAPI	Vector Laboratories	Cat#H-1200
Ubiquitin-activating enzyme E1 (UBE1A)	Boston Biochem	Cat#E-305
Ubiquitin-conjugating enzyme E2 (UBE2L3)	Boston Biochem	Cat#E2-640
Ubiquitin	Boston Biochem	Cat#U-100
Methyl-ubiquitin	Boston Biochem	Cat#U-501
K6-only ubiquitin	Boston Biochem	Cat#UM-K60
K11-only ubiquitin	Boston Biochem	Cat#UM-K110
K27-only ubiquitin	Boston Biochem	Cat#UM-K270
K29-only ubiquitin	Boston Biochem	Cat#UM-K290
K33-only ubiquitin	Boston Biochem	Cat#UM-K330
K48-only ubiquitin	Boston Biochem	Cat#UM-K480
K63-only ubiquitin	Boston Biochem	Cat#UM-K630
Histone H2A (1-22) - GK(Biotin)	AnaSpec	Cat#64639-1
Histone H3 (1-21) Biotinylated	AnaSpec	Cat#AS-61702
Histone H4 (1-23) - GGK(Biotin)	AnaSpec	Cat#AS-65097
KOD DNA polymerase	Merck Millipore	Cat#71086-4
Phusion DNA polymerase	NEB	Cat#M0530L
Critical Commercial Assays
Dual-Luciferase Reporter Assay System	Promega	Cat#E1910
UbiCREST Deubiquitinase Enzyme Kit	Boston Biochem	Cat#K-400
RNeasy Mini Kit (RNA Purification)	QIAGEN	Cat#74104
iScript cDNA synthesis kit	Biorad	Cat#170-8890
SYBR Select Master Mix	Applied Biosystems	Cat#4472908
Deposited Data
Raw Imaging Data	This paper	http://dx.doi.org/10.17632/j2wjdkj5cn.1
Experimental Models: Cell Lines
HEK293T	ATCC	Cat#CRL-3216
HeLa	ATCC	Cat#CCL-2
HCT116	ATCC	Cat#CCL-247
Experimental Models: Organisms/Strains
D. melanogaster: hydK7-19	Prof. Jessica E. Treisman	FlyBase: FBal0144234
D. melanogaster: groMB36	Prof. David Ish-Horowicz	FlyBase: FBal0230454
D. melanogaster: axinP	Prof. Tetsu Akiyama	FlyBase: FBal0097414
D. melanogaster: UAS.SoxF	Dr. Fernando Casares	FlyBase: FBtp0051564
D. melanogaster: pygoS123	Bloomington Drosophila Stock Center	FlyBase: FBal0146872 Bloomington: 7209
D. melanogaster: UAS.ArmS10	Bloomington Drosophila Stock Center	FlyBase: FBtp0001723 Bloomington: 4782
D. melanogaster: UAS.MamDN	Prof. Sarah Bray	FlyBase: FBtp0014588
D. melanogaster: Vg-Gal4, UAS-flp; FRT82b GFP	Bienz Laboratory	Miller et al., 2013
Oligonucleotides
Primer sequences for RT-qPCR, see Table S3	This paper	N/A
Recombinant DNA
Plasmid: pCMV-tag2b-Flag-UBR5	Prof. Rina Rosin-Arbesfeld	Hay-Koren et al., 2011
Plasmid: pCS2 Myc-TLE3	Prof. Ethan Lee	Hanson et al., 2012
Plasmid: pcDNA3.1 VCP-GFP	Prof. Nico Dantuma	Tresse et al., 2010
Plasmid: pcDNA3.1 HA-Δ45-β-catenin	Bienz Laboratory	Morin et al., 1997
Plasmid: pET GST-HES1	Prof. Stefano Stifani	Grbavec and Stifani, 1996
Plasmid: pCMV His-ubiquitin	Dr. Thomas Mund	Mund et al., 2015
Plasmid: pcDNA3.1 HA-PAIP2	Bienz Laboratory	N/A
Plasmid: pcDNA3.1 SSDP-GFP	Bienz Laboratory	Fiedler et al., 2015
Plasmid: pcDNA3.1 LDB1-GFP	Bienz Labaratory	Fiedler et al., 2015
Plasmid: pcDNA3.1 GFP-TCF4	Bienz Labaratory	N/A
Plasmid: pcDNA3.1 GFP-Pygo1	Bienz Labaratory	N/A
Plasmid: pSpCas9(BB)-2A-GFP (PX458)	Addgene	Ran et al., 2013; Addgene#48138
Plasmid: pSpCas9(BB)-2A-Puro (PX459)	Addgene	Ran et al., 2013; Addgene#62988
Plasmid: pTA-Luc m50 Super 8x TopFLASH	Addgene	Veeman et al., 2003; Addgene#12456
Plasmid: pRL-CMV Renilla luciferase	Promega	Cat#E2261