Abstract
Beta-transducin repeat containing protein 1 (β-TrCP1) is a versatile F-box protein that is responsible for substrate recognition of SCFβ-TrCP1 ubiquitin ligase. In human cells, two major alternatively spliced isoforms (b and f) of β-TrCP1 were found. Recently, we identified that CENP-W interacts with the β-TrCP1 and regulates the cellular distribution of β-TrCP1. In this study, we examined whether CENP-W, a new kinetochore component, may differentially regulate the two major isoforms of human β-TrCP1 (b and f), especially in the cytoplasmic-nuclear shuttling of β-TrCP1. An in vivo binding assay was performed to examine whether CENP-W binds differently to the two isoforms of β-TrCP1. EGFP-conjugated β-TrCP1 isoforms were co-transfected with NLS-defective mutant CENP-W and their cellular distribution were observed using a fluorescence microscopy. Although CENP-W interacts with both b and f isoforms, it has a greater affinity for the b isoform rather than f isoform. Moreover, CENP-W effectively regulates the nuclear-cytoplasmic shuttling of these two β-TrCP1 isoforms, but with a slight preference towards the b isoform. The Elongin C-binding motif existing in the b isoform may be involved in their specific association. CENP-W showed a higher affinity toward the β-TrCP1 b isoform, and translocated isoform b more efficiently than isoform f, which may allow a fine regulation of of β-TrCP1 in the cells.
Similar content being viewed by others
Abbreviations
- SCF:
-
SKP1-Cullin 1-F-box protein
- β-TrCP:
-
Beta-transducin repeat containing protein
- CRL:
-
Cullin-RING ubiquitin ligase
- VHL:
-
Von Hippel–Lindau
- CENP-W:
-
Centromere protein W
- NLS:
-
Nuclear localization signal
References
Ang XL, Wade Harper J (2005) SCF-mediated protein degradation and cell cycle control. Oncogene 24:2860–2870
Besnard-Guerin C, Belaidouni N, Lassot I, Segeral E, Jobart A, Marchal C, Benarous R (2004) HIV-1 Vpu sequesters beta-transducin repeat-containing protein (betaTrCP) in the cytoplasm and provokes the accumulation of beta-catenin and other SCFbetaTrCP substrates. J Biol Chem 279:788–795
Cheon Y, Jeon S, Lee S (2016) Centromere protein W interacts with beta-transducin repeat-containing protein 1 and modulates its subcellular localization. FEBS Lett 590:4441–4452
Chun Y, Park B, Koh W, Lee S, Cheon Y, Kim R, Che L (2011) New centromeric component CENP-W is an RNA-associated nuclear matrix protein that interacts with nucleophosmin/B23 protein. J Biol Chem 286:42758–42769
Chun Y, Lee M, Park B, Lee S (2013) CSN5/JAB1 interacts with the centromeric components CENP-T and CENP-W and regulates their proteasome-mediated degradation. J Biol Chem 288:27208–27219
Chun Y, Kim R, Lee S (2016) Centromere protein (CENP)-W interacts with heterogeneous nuclear ribonucleoprotein (hnRNP) U and may contribute to kinetochore-microtubule attachment in mitotic cells. PLoS ONE 11:e0149127
Davis M, Hatzubai A, Andersen JS, Ben-Shushan E, Fisher GZ, Yaron A, Bauskin A, Mercurio F, Mann M, Ben-Neriah Y (2002) Pseudosubstrate regulation of the SCF(beta-TrCP) ubiquitin ligase by hnRNP-U. Genes Dev 16:439–451
Frescas D, Pagano M (2008) Deregulated proteolysis by the F-box proteins SKP2 and beta-TrCP: tipping the scales of cancer. Nat Rev Cancer 8:438–449
Fuchs SY, Spiegelman VS, Kumar KG (2004) The many faces of beta-TrCP E3 ubiquitin ligases: reflections in the magic mirror of cancer. Oncogene 23:2028–2036
Jin J, Shirogane T, Xu L, Nalepa G, Qin J, Elledge SJ, Harper JW (2003) SCFbeta-TRCP links Chk1 signaling to degradation of the Cdc25A protein phosphatase. Genes Dev 17:3062–3074
Jin J, Cardozo T, Lovering RC, Elledge SJ, Pagano M, Harper JW (2004) Systematic analysis and nomenclature of mammalian F-box proteins. Genes Dev 18:2573–2580
Kaczmarczyk A, Sullivan KF (2014) CENP-W plays a role in maintaining bipolar spindle structure. PLoS ONE 9:e106464
Kamura T, Sato S, Haque D, Liu L, Kaelin WG Jr, Conaway RC, Conaway JW (1998) The Elongin BC complex interacts with the conserved SOCS-box motif present in members of the SOCS, ras, WD-40 repeat, and ankyrin repeat families. Genes Dev 12:3872–3881
Kim H, Lee M, Lee S, Park B, Koh W, Lee DJ, Lim DS (2009) Cancer-upregulated gene 2 (CUG2), a new component of centromere complex, is required for kinetochore function. Mol Cells 27:697–701
Koh W, Park B, Lee S (2015) A new kinetochore component CENP-W interacts with the polycomb-group protein EZH2 to promote gene silencing. Biochem Biophys Res Commun 464:256–262
Lee S, Gang J, Jeon SB, Choo SH, Lee B, Kim YG, Lee YS, Jung J, Song SY, Koh SS (2007) Molecular cloning and functional analysis of a novel oncogene, cancer-upregulated gene 2 (CUG2). Biochem Biophys Res Commun 360:633–639
Margottin F, Bour SP, Durand H, Selig L, Benichou S, Richard V, Thomas D, Strebel K, Benarous R (1998) A novel human WD protein, h-beta TrCp, that interacts with HIV-1 Vpu connects CD4 to the ER degradation pathway through an F-box motif. Mol Cell 1:565–574
Nakayama KI, Nakayama K (2006) Ubiquitin ligases: cell-cycle control and cancer. Nat Rev Cancer 6:369–381
Okumura F, Matsuzaki M, Nakatsukasa K, Kamura T (2012) The role of elongin BC-containing ubiquitin ligases. Front Oncol 2:10
Ougolkov A, Zhang B, Yamashita K, Bilim V, Mai M, Fuchs SY, Minamoto T (2004) Associations among beta-TrCP, an E3 ubiquitin ligase receptor, beta-catenin, and NF-kappaB in colorectal cancer. J Natl Cancer Inst 96:1161–1170
Porkka K, Saramaki O, Tanner M, Visakorpi T (2002) Amplification and overexpression of elongin C gene discovered in prostate cancer by cDNA microarrays. Lab Invest 82:629–637
Putters J, Slotman JA, Gerlach JP, Strous GJ (2011) Specificity, location and function of betaTrCP isoforms and their splice variants. Cell Signal 23:641–647
Seo E, Kim H, Kim R, Yun S, Kim M, Han JK, Costantini F, Jho EH (2009) Multiple isoforms of beta-TrCP display differential activities in the regulation of Wnt signaling. Cell Signal 21:43–51
Suzuki H, Chiba T, Kobayashi M, Takeuchi M, Suzuki T, Ichiyama A, Ikenoue T, Omata M, Furuichi K, Tanaka K (1999) IkappaBalpha ubiquitination is catalyzed by an SCF-like complex containing Skp1, cullin-1, and two F-box/WD40-repeat proteins, betaTrCP1 and betaTrCP2. Biochem Biophys Res Commun 256:127–132
Wu G, Xu G, Schulman BA, Jeffrey PD, Harper JW, Pavletich NP (2003a) Structure of a beta-TrCP1-Skp1-beta-catenin complex: destruction motif binding and lysine specificity of the SCF (beta-TrCP1) ubiquitin ligase. Mol Cell 11:1445–1456
Wu G, Xu G, Schulman BA, Jeffrey PD, Harper JW, Pavletich NP (2003b) Structure of a β-TrCP1-Skp1-β-catenin complex. Mol Cell 11:1445–1456
Acknowledgements
This work was supported by a research fund from Chungnam National University.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
Yeongmi Cheon and Soojin Lee declare that they have no conflicts of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Rights and permissions
About this article
Cite this article
Cheon, Y., Lee, S. Two major alternative splice variants of beta-TrCP1 interact with CENP-W with different binding preferences. Genes Genom 41, 167–174 (2019). https://doi.org/10.1007/s13258-018-0748-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13258-018-0748-3