Abstract
Mutational inactivation of adenomatous polyposis coli (APC) initiates most colon carcinomas. APC functions include targeting cytoplasmic beta-catenin, a Wnt pathway mediator, for proteolysis. Although APC shuttles between cytoplasm and nucleus, the role of nuclear APC protein, particularly with respect to nuclear beta-catenin levels and activity, remains unclear. Here, we demonstrate that APC lacking functional nuclear localization signals (NLSs) or nuclear export signals (NESs) does not effectively downregulate nuclear beta-catenin levels; neither does wild-type APC when nuclear export is blocked. While APC bearing mutated NLSs could not downregulate beta-catenin-mediated transcriptional activation, APC lacking NESs remained active. Consistent with the hypothesis that nuclear APC lacking NESs can inhibit beta-catenin function by sequestration, we show that endogenous APC and beta-catenin proteins interact within the nucleus. These data demonstrate that nuclear APC binding to beta-catenin, and then inducing its nuclear export, plays a critical role in the control of nuclear beta-catenin levels and activity.
MeSH Terms
Adenomatous Polyposis Coli/genetics,metabolism
Antibiotics, Antineoplastic/pharmacology
Blotting, Western
Cell Line
Cell Nucleus/metabolism
Cloning, Molecular
Colonic Neoplasms/genetics
Cytoplasm/metabolism
Cytoskeletal Proteins/biosynthesis,genetics
DNA-Binding Proteins/metabolism
Down-Regulation
Fatty Acids, Unsaturated/pharmacology
Humans
Lymphoid Enhancer-Binding Factor 1
Microscopy, Fluorescence
Models, Genetic
Mutation
Precipitin Tests
Protein Binding
Protein Transport
Trans-Activators
Transcription Factors/metabolism
Transcriptional Activation
Transfection
beta Catenin
Chemicals
Antibiotics, Antineoplastic
CTNNB1 protein, human
Cytoskeletal Proteins
DNA-Binding Proteins
Fatty Acids, Unsaturated
Lymphoid Enhancer-Binding Factor 1
Trans-Activators
Transcription Factors
beta Catenin
leptomycin B
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Neufeld K L
Department of Oncological Sciences, Huntsman Cancer Institute, University of Utah, Salt Lake City 84112, USA. kristi.neufeld@genetics.utah.edu
Zhang F
Cullen B R
White R L
References (19)
19 references, click to expand
-
The APC tumour suppressor has a nuclear export function.
Nature. 2000 Aug 31;406(6799):1009-12
PMID: 10984057
-
Membrane-anchored plakoglobins have multiple mechanisms of action in Wnt signaling.
Mol Biol Cell. 1999 Oct;10(10):3151-69
PMID: 10512857
-
Adenomatous polyposis coli protein contains two nuclear export signals and shuttles between the nucleus and cytoplasm.
Proc Natl Acad Sci U S A. 2000 Oct 24;97(22):12085-90
PMID: 11035805
-
Phosphorylation near nuclear localization signal regulates nuclear import of adenomatous polyposis coli protein.
Proc Natl Acad Sci U S A. 2000 Nov 7;97(23):12577-82
PMID: 11050185
-
APC mutations occur early during colorectal tumorigenesis.
Nature. 1992 Sep 17;359(6392):235-7
PMID: 1528264
-
Regulation of intracellular beta-catenin levels by the adenomatous polyposis coli (APC) tumor-suppressor protein.
Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):3046-50
PMID: 7708772
-
Identification of a signal for rapid export of proteins from the nucleus.
Cell. 1995 Aug 11;82(3):463-73
PMID: 7634336
-
Binding of GSK3beta to the APC-beta-catenin complex and regulation of complex assembly.
Science. 1996 May 17;272(5264):1023-6
PMID: 8638126
-
Forced expression of the tumor suppressor adenomatosis polyposis coli protein induces disordered cell migration in the intestinal epithelium.
Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9588-93
PMID: 8790374
-
Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF.
Cell. 1997 Mar 21;88(6):789-99
PMID: 9118222
-
Nuclear and cytoplasmic localizations of the adenomatous polyposis coli protein.
Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3034-9
PMID: 9096341
-
Loss of beta-catenin regulation by the APC tumor suppressor protein correlates with loss of structure due to common somatic mutations of the gene.
Cancer Res. 1997 Oct 15;57(20):4624-30
PMID: 9377578
-
Identification of c-MYC as a target of the APC pathway.
Science. 1998 Sep 4;281(5382):1509-12
PMID: 9727977
-
E-cadherin binding prevents beta-catenin nuclear localization and beta-catenin/LEF-1-mediated transactivation.
J Cell Sci. 1999 Apr;112 ( Pt 8):1237-45
PMID: 10085258
-
Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells.
Nature. 1999 Apr 1;398(6726):422-6
PMID: 10201372
-
In or out? Regulating nuclear transport.
Curr Opin Cell Biol. 1999 Apr;11(2):241-7
PMID: 10209150
-
Nuclear localization and formation of beta-catenin-lymphoid enhancer factor 1 complexes are not sufficient for activation of gene expression.
Mol Cell Biol. 1999 Jun;19(6):4503-15
PMID: 10330189
-
The ubiquitin-proteasome pathway and serine kinase activity modulate adenomatous polyposis coli protein-mediated regulation of beta-catenin-lymphocyte enhancer-binding factor signaling.
J Biol Chem. 1999 Jun 4;274(23):16641-5
PMID: 10347231
-
Nuclear-cytoplasmic shuttling of APC regulates beta-catenin subcellular localization and turnover.
Nat Cell Biol. 2000 Sep;2(9):653-60
PMID: 10980707