Abstract
The E2 polyubiquitin-conjugating enzyme Ubc13 is a mediator of innate immune reactions. Ubc13 mediates the conjugation of keratin (K)63-linked polyubiquitin chains onto TNF receptor-associated factor 6 and IKKγ during NF-κB activation. In contrast to K48-linked polyubiquitin chains, K63-linked polyubiquitin chains function in nonproteasomal biological processes. Although Ubc13 has been shown to be critical for Toll-like receptor (TLR) and IL-1 receptor signaling, the function of Ubc13 in the epidermis has not been studied. We generated keratinocyte-specific Ubc13-deficient mice (Ubc13(flox/flox)K5-Cre). At birth, the skin of the Ubc13(flox/flox)K5-Cre mice was abnormally shiny and smooth; in addition, the mice did not grow and died by postnatal day 2. Histological analysis showed atrophy of the epidermis with keratinocyte apoptosis. Immunohistochemical analyses revealed reduced proliferation, abnormal differentiation, and apoptosis of keratinocytes in the Ubc13(flox/flox)K5-Cre mouse epidermis. In culture, Ubc13(flox/flox)K5-Cre keratinocyte growth was impaired, and spontaneous cell death occurred. Moreover, the deletion of Ubc13 from cultured Ubc13(flox/flox) keratinocytes by means of an adenoviral vector carrying Cre recombinase also resulted in spontaneous cell death. Therefore, Ubc13 is essential for keratinocyte growth, differentiation, and survival. Analyses of intracellular signaling revealed that the IL-1 and TNF-induced activation of JNK, p38, and NF-κB pathways was impaired in Ubc13(flox/flox)K5-Cre keratinocytes. In conclusion, Ubc13 appears to be essential for epidermal integrity in mice.
MeSH Terms
Animals
Cell Differentiation/physiology
Cell Survival/physiology
Epidermis/immunology,metabolism
I-kappa B Kinase/genetics,metabolism
Immunity, Innate/physiology
Keratinocytes/immunology,metabolism
Keratins/genetics,immunology,metabolism
MAP Kinase Signaling System/physiology
Mice
Mice, Knockout
Receptors, Interleukin-1/genetics,immunology,metabolism
TNF Receptor-Associated Factor 6/genetics,immunology,metabolism
Toll-Like Receptors/genetics,immunology,metabolism
Ubiquitin-Conjugating Enzymes/genetics,immunology,metabolism
Ubiquitination/physiology
Chemicals
Receptors, Interleukin-1
TNF Receptor-Associated Factor 6
Toll-Like Receptors
Keratins
Ube2n protein, mouse
Ubiquitin-Conjugating Enzymes
I-kappa B Kinase
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Sayama Koji
Department of Dermatology, Ehime University Graduate School of Medicine, Ehime 791-0295, Japan. sayama@m.ehime-u.ac.jp
Yamamoto Masahiro
Shirakata Yuji
Hanakawa Yasushi
Hirakawa Satoshi
Dai Xiuju
Tohyama Mikiko
Tokumaru Sho
Shin Myoung-Sook
Sakurai Hiroaki
Akira Shizuo
Hashimoto Koji
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