Kyushu University Academic Staff Educational and Research Activities Database
Researcher information (To researchers) Need Help? How to update
Keiichi Nakayama Last modified date:2017.07.12

Graduate School

Academic Degree
M.D., Ph.D.
Field of Specialization
Proteolysis to regulate cell cycle
Outline Activities
Research Project:Investigation into the molecular mechanism of cell-growth and cell-death control.

1. Elucidation of physiological function of p27Kip1 that controls cell growth and differentiation

Growth and death are the basic properties of the cell. Disturbance of the control for cell growth and death has been believed as a major cause of carcinogenesis. Cell growth and death do not always take place, rather they are tightly regulated in tissue- and development-specific manners. For example, neurons proliferate only in fetus, but never in adult. The mechanism of this development-specific regulation of cell growth has been largely unclear.
We investigate the physiological roles of factors which control cell cycle and cell death. Recently we focus our interest on the roles of cyclin-dependent-kinase inhibitors (CKI), which negatively regulate cell proliferation, in development and carcinogenesis. Mice lacking p27Kip1, which is the most related CKI to development and growth regulation, were created in our laboratory. The p27Kip1 knockout mice displayed increased body size, and multiple organ hyperplasia in immune, nervous, and reproductive systems. In addition, pituitary tumor frequently arose in these knockout mice, indicating that p27Kip1 is an oncosuppressor molecule. A series of clinical studies suggested that tumors with lower expression of p27Kip1 have poorer prognosis. Thus, we explore the molecular mechanism by which expression level of p27Kip1 is controlled.

2. Elucidation of the molecular mechanism for ubiquitination of p27Kip1

The ubiquitin-proteasome pathway of protein degradation plays an important role in control of the abundance of short-lived regulatory proteins. The ubiquitin-attachment system consists of several components that act in concert. A ubiquitin-activating enzyme (E1) uses ATP to form a thioester bond between itself and ubiquitin, and it then transfers the activated ubiquitin to a ubiquitin-conjugating enzyme (E2). Protein-ubiquitin ligation often requires the participation of a third component, termed ubiquitin ligase (E3). Although the E3 components are thought to be primarily responsible for substrate recognition, they are the least well understood of the enzymes of the ubiquitin-conjugation system.
The SCF complexes, a major class of E3 ligases, consist of the invariable components Skp1, Cul1, and Rbx1/ROC1 as well as a variable component, known as an F-box protein, that binds to Skp1 through its F-box motif and serves as the substrate-recognition subunit. Skp2, an F-box protein with leucine-rich repeats, mediates degradation of p27Kip1.
With the use of gene targeting in embryonic stem (ES) cells, we have now generated mice lacking Skp2. Cells derived from these animals exhibit hyperaccumulation of both cyclin E and p27Kip1, polyploidy, and multiple centrosomes. Consistent with these observations, our biochemical analysis indicates that Skp2 mediates ubiquitin-dependent degradation of cyclin E. Taken together, these data suggest that SCFSkp2 functions as the principal ubiquitin ligase in determining the abundance of cell cycle regulatory proteins at the G1-S transition, thereby ensuring strict control of chromosomal replication and centrosome duplication.
Research Interests
  • Elucidation of mechanisms underlying proteolysis in the cell cycle control
    keyword : cell cycle, p27, ubiquitin, knockout mouse
    1996.10We have been studying the mechanisms underlying proteolysis of p27 CDK inhibitor. We have created a series of mutant mice lacking the key molecules that are involved in the p27 degradation..
  • Study of ubiquitylation in neurodegerative diseases
    keyword : polyglutamine disease, SCA3, ataxin-3, ubiquitin, E4B/UFD2a
    2001.04Various inherited neurodegenerative diseases result from an increase in the number of glutamine codon repeats within the open reading frame of the responsible gene. The presence of insoluble aggregates in neurons, which have been shown to stain with anti-ubiquitin, is a hallmark of these polyglutamine diseases as well as of many other neurodegenerative disorders. However, neither the significance of these inclusions nor the molecular mechanism of their ubiquitylation has been clear. We have now shown that ataxin-3, the product of the gene responsible for a polyglutamine disease Spinocerebellar ataxia 3 (SCA3), is degraded by the ubiquitin-proteasome pathway. To identify the enzymes that mediate the ubiquitylation of ataxin-3, we partially purified a polyubiquitylation activity for this protein, and found that E4B (a ubiquitin chain assembly factor, or E4) as well as VCP (a AAA-family ATPase) copurified with this polyubiquitylation activity. Significantly, ataxin-3-VCP-E4B formed a trimeric complex, and E4B mediated polyubiquitylation of ataxin-3. Expression of E4B markedly promoted the degradation of an ataxin-3 isoform with an expanded polyglutamine tract, overcoming the resistance to degradation conferred by glutamine repeat expansion. In contrast, functional inhibition of E4B resulted both in a delay in ataxin-3 degradation and in the consequent formation of intracellular aggregates. Finally, we showed that expression of E4B prevented neurodegeneration in transgenic flies expressing an ataxin-3 mutant with an expanded polyglutamine tract. Our data thus provide biochemical and genetic evidence that a mammalian E4 enzyme plays an important role in the degradation of ataxin-3. Our study is one of a few that have recently unveiled the mechanisms of ubiquitylation of proteins involved in neurodegenerative diseases, and it represents the first demonstration that E4 exists in multicellular organisms and contributes to the degradation of a physiological substrate. Targeted expression of E4B is therefore a potential gene therapy for polyglutamine diseases..
Current and Past Project
  • Molecular analysis of the SCF ubiquitin ligase from yeast to human
Academic Activities
1. Nakayama, K. I., Nakayama, K., Ubiquitin ligases: cell-cycle control and cancer, Nature Rev. Cancer, 2006.05.
1. Keiichi Nakayama, Robotic crowd biology with Maholo LabDroids, 2017.07.
2. Keiichi Nakayama, Hippo signaling suppresses cell ploidy and tumorigenesis through Skp2, CANCER CELL, 31, 5, 669-+, 2017.05.
3. Keiichi Nakayama, FBXL5 inactivation in mouse brain induces aberrant proliferation of neural stem progenitor cells, MOLECULAR AND CELLULAR BIOLOGY, 37, 8, 2017.04.
4. Keiichi Nakayama, A large-scale targeted proteomics assay resource based on an in vitro human proteome, NATURE METHODS, 14, 3, 251-+, 2017.03.
5. Keiichi Nakayama, mTORC1 and muscle regeneration are regulated by the LINC00961-encoded SPAR polypeptide, NATURE, 541, 7636, 228-+, 2017.01.
6. Keiichi Nakayama, SRRM4-dependent neuron-specific alternative splicing of protrudin transcripts regulates neurite outgrowth, SCIENTIFIC REPORTS, 7, 2017.01.
7. Keiichi Nakayama, Phosphoproteomics analyses show subnetwork systems in T-cell receptor signaling, GENES TO CELLS, 21, 10, 1095-1112, 2016.10.
8. Keiichi Nakayama, CHD8 haploinsufficiency results in autistic-like phenotypes in mice, NATURE, 537, 7622, 675-+, 2016.09.
9. Keiichi Nakayama, FBXL12 regulates T-cell differentiation in a cell-autonomous manner, STEM CELLS, 21, 5, 517-524, 2016.05.
10. Keiichi Nakayama, FBXL12-mediated degradation of ALDH3 is essential for trophoblast differentiation during placental development, STEM CELLS, 33, 11, 2015.11.
11. Keiichi Nakayama, Maternal TET3 is dispensable for embryonic development but is required for neonatal growth, SCIENTIFIC REPORTS, 5, 2015.09.
12. Keiichi Nakayama, F-box protein FBXW7 inhibits-cancer metastasis in a non-cell-autonomous manner, JOURNAL OF CLINICAL INVESTIGATION, 125, 2, 621-635, 2015.02.
13. Matsumoto Akinobu, Shoichiro Takeishi, Keiichi Nakayama, p57 regulates T-cell development and prevents lymphomagenesis by balancing p53 activity and pre-TCR signaling, BLOOD, 123, 22, 2014.05.
14. Linsey Reavie, Shannon M. Buckley, Evangelia Loizou, Shoichiro Takeishi, Beatriz Aranda-Orgilles, Delphine Ndiaye-Lobry, Omar Abdel-Wahab, Sherif Ibrahim, Keiichi Nakayama, Iannis Aifantis, Regulation of c-Myc Ubiquitination Controls Chronic Myelogenous Leukemia Initiation and Progression, CANCER CELL, 23, 3, 362-375, 2013.03.
15. Shoichiro Takeishi, Matsumoto Akinobu, Ichiro Onoyama, Ichiro Naka, Atsushi Hirao, Keiichi Nakayama, Ablation of Fbxw7 Eliminates Leukemia-Initiating Cells by Preventing Quiescence, CANCER CELL, 23, 3, 347-361, 2013.03.
16. Arisa Hirano, Kanae Yumimoto, Ryosuke Tsunematsu, Masaki Matsumoto, Masaaki Oyama, Hiroko Kozuka-Hata, Tomoki Nakagawa, Darin Lanjakornsiripan, Keiichi Nakayama, Yoshitaka Fukada, FBXL21 Regulates Oscillation of the Circadian Clock through Ubiquitination and Stabilization of Cryptochromes, CELL, 152, 5, 1106-1118, 2013.02.
17. Shotaro Saita, Michiko Shirane, Keiichi Nakayama, Selective escape of proteins from the mitochondria during mitophagy, NATURE COMMUNICATIONS, 4, 2013.01.
18. Yasutaka Okita, Keiichi Nakayama, UPS Delivers Pluripotency, CELL STEM CELL, 11, 6, 728-730, 2012.12.
19. Hidefumi Fukushima, Matsumoto Akinobu, Hiroyuki Inuzuka, Bo Zhai, Alan W. Lau, Lixin Wan, Daming Gao, Shavali Shaik, Min Yuan, Steven P. Gygi, Eijiro Jimi, John M. Asara, Keiko Nakayama, Wenyi Wei, Keiichi Nakayama, SCFFbw7 Modulates the NF kappa B Signaling Pathway by Targeting NF kappa B2 for Ubiquitination and Destruction, CELL REPORTS, 1, 5, 434-443, 2012.05.
20. Chia-Hsin Chan, Chien-Feng Li, Wei-Lei Yang, Yuan Gao, Szu-Wei Lee, Zizhen Feng, Hsuan-Ying Huang, Leo G. Flores, Kelvin K.C. Tsai, Yiping Shao, John D. Hazle, Dihua Yu, Wenyi Wei, Dos Sarbassov, Mien-Chie Hung, Keiichi Nakayama, Hui-Kuan Lin, The Skp2-SCF E3 Ligase Regulates Akt Ubiquitination, Glycolysis, Herceptin Sensitivity, and Tumorigenesis, CELL, 149, 5, 1098-1111, 2012.05.
21. Moroishi, T., Nishiyama, M., Takeda, Y., Iwai, K., Nakayama, K.I., The FBXL5-IRP2 axis is integral to control of iron metabolism in vivo, Cell Metabolism, 14: 339-51, 2011.09.
22. Matsumoto, A., Takeishi, S., Kanie, T., Susaki, E., Onoyama, I., Tateishi, Y., Nakayama, K., Nakayama, K.I., p57 is required for quiescence and maintenance of adult hematopoietic stem cells, Cell Stem Cell, 9: 262-71, 2011.09.
23. Onoyama, I., Suzuki, A., Matsumoto, A., Tomita, K., Katagiri, H., Oike, Y., Nakayama, K., Nakayama, K.I., Fbxw7 regulates lipid metabolism and cell fate decisions in the mouse liver, J. Clin. Invest., 121, 342-354, 2011.01.
24. Liu Z, Liu X, Nakayama KI, Nakayama K, Ye K., Protein kinase C-delta phosphorylates Ebp1 and prevents its proteolytic degradation, enhancing cell survival., J Neurochem., 100(5):1278-88., 2007.03.
25. Moller C, Karlberg M, Abrink M, Nakayama KI, Motoyama N, Nilsson G., Bcl-2 and Bcl-XL are indispensable for the late phase of mast cell development from mouse embryonic stem cells., Exp Hematol., 35(3):385-93., 2007.03.
26. Tu X, Joeng KS, Nakayama KI, Nakayama K, Rajagopal J, Carroll TJ, McMahon A., Noncanonical Wnt signaling through G protein-linked PKCdelta activation promotes bone formation., Dev Cell. , 12(1):113-27., 2007.01.
27. Uchida T, Iwashita N, Ohara-Imaizumi M, Ogihara T, Nagai S, Choi JB, Tamura Y, Tada N, Kawamori R, Nakayama KI, Nagamatsu S, Watada H., Protein kinase Cdelta plays a non-redundant role in insulin secretion in pancreatic beta cells., J Biol Chem., 282(4):2707-16., 2007.01.
28. Shukla A, Lounsbury KM, Barrett TF, Gell J, Rincon M, Butnor KJ, Taatjes DJ, Davis GS, Vacek P, Nakayama KI, Nakayama K, Steele C, Mossman BT., Asbestos-induced peribronchiolar cell proliferation and cytokine production are attenuated in lungs of protein kinase C-knockout mice., J Biol Chem., 170(1):140-51., 2007.01.
29. Sakai T, Sakaue H, Nakamura T, Okada M, Matsuki Y, Watanabe E, Hiramatsu R, Nakayama K, Nakayama KI, Kasuga M., Skp2 controls adipocyte proliferation during the development of obesity., J Biol Chem., 282(3):2038-46., 2007.01.
30. Yanagawa M, Tsukuba T, Nishioku T, Okamoto Y, Okamoto K, Takii R, Terada Y, Nakayama KI, Kadowaki T, Yamamoto K., Cathepsin E deficiency induces a novel form of lysosomal storage disorder showing the accumulation of lysosomal membrane sialoglycoproteins and the elevation of lysosomal pH in macrophages., J Biol Chem., 82(3):1851-62., 2007.01.
31. Itoh Y, Masuyama N, Nakayama K, Nakayama KI, Gotoh Y., The cyclin-dependent kinase inhibitors p57 and p27 regulate neuronal migration in the developing mouse neocortex., J Biol Chem., 282(1):390-6., 2007.01.
32. Shirane M, Nakayama KI., Protrudin induces neurite formation by directional membrane trafficking., Science., 314(5800):818-21., 2006.11.
33. Matsumoto A, Onoyama I, Nakayama KI., Expression of mouse Fbxw7 isoforms is regulated in a cell cycle- or p53-dependent manner., Biochem Biophys Res Commun., 3350(1):114-9., 2006.11.
34. Hara K, Nakayama KI, Nakayama K., Geminin is essential for the development of preimplantation mouse embryos., Genes Cells., 11(11):1281-93., 2006.11.
35. Tsunematsu R, Nishiyama M, Kotoshiba S, Saiga T, Kamura T, Nakayama KI., Fbxw8 is essential for Cul1-Cul7 complex formation and for placental development., Mol Cell Biol., 26(16):6157-69., 2006.10.
36. Fujii Y, Yada M, Nishiyama M, Kamura T, Takahashi H, Tsunematsu R, Susaki E, Nakagawa T, Matsumoto A, Nakayama KI., Fbxw7 contributes to tumor suppression by targeting multiple proteins for ubiquitin-dependent degradation., Cancer Sci., 97(8):729-36., 2006.10.
37. Parcellier A, Brunet M, Schmitt E, Col E, Didelot C, Hammann A, Nakayama K, Nakayama KI, Khochbin S, Solary E, Garrido C., HSP27 favors ubiquitination and proteasomal degradation of p27Kip1 and helps S-phase re-entry in stressed cells., FASEB J., 20(8):1179-81., 2006.06.
38. Yoshida K, Yamaguchi T, Shinagawa H, Taira N, Nakayama KI, Miki Y., Protein kinase C delta activates topoisomerase IIalpha to induce apoptotic cell death in response to DNA damage., Mol Cell Biol., 26(9):3414-31., 2006.06.
39. Fotovati A, Nakayama K, Nakayama KI., Impaired germ cell development due to compromised cell cycle progression in Skp2-deficient mice., Cell Div., 1:4., 2006.04.
40. Humphries MJ, Limesand KH, Schneider JC, Nakayama KI, Anderson SM, Reyland ME., Suppression of apoptosis in the protein kinase Cdelta null mouse in vivo., J Biol Chem., 281(14):9728-37., 2006.04.
41. Kase S, Yoshida K, Ohgami K, Shiratori K, Ohno S, Nakayama KI., phorylation of p27(KIP1) in the mitotic cells of the corneal epithelium., Curr Eye Res., 31(4):307-12., 2006.04.
42. Nishitani H, Sugimoto N, Roukos V, Nakanishi Y, Saijo M, Obuse C, Tsurimoto T, Nakayama KI, Nakayama K, Fujita M, Lygerou Z, Nishimoto T., Two E3 ubiquitin ligases, SCF-Skp2 and DDB1-Cul4, target human Cdt1 for proteolysis., EMBO J., 25(5):1126-36., 2006.03.
43. Kase S, Yoshida K, Ohgami K, Shiratori K, Suzuki Y, Nakayama KI, Ohno S., Expression of cdc2 and p27(KIP1) phosphorylation in mitotic cells of the human retinoblastoma., Int J Mol Med., 17(3):465-8., 2006.03.
44. Kase S, Yoshida K, Harada T, Harada C, Namekata K, Suzuki Y, Ohgami K, Shiratori K, Nakayama KI, Ohno S., Phosphorylation of extracellular signal-regulated kinase and p27(KIP1) after retinal detachment., Graefes Arch Clin Exp Ophthalmol., 244(3):352-8, 2006.03.
45. Shirane M, Nakayama KI., Inherent calcineurin inhibitor FKBP38 targets Bcl-2 to mitochondria and inhibits apoptosis., Nature Cell Biol., 5, 1, 5(1): 28-37, 2003.01.
Other Educational Activities
  • 2017.04.
  • 2016.04.
  • 2015.05.
  • 2015.04.
  • 2014.04.
  • 2013.04.
  • 2012.04.
  • 2011.04.
  • 2010.04.
  • 2009.04.
  • 2008.05.
  • 2007.05.
  • 2006.05.
  • 2005.05.
  • 2004.05.
  • 2003.05.
  • 2002.05.
  • 2001.05.
  • 2000.05.
  • 1999.05.
  • 1998.05.