Pin1 associates with and induces translocation of CRTC2 to the cytosol, thereby suppressing cAMP-responsive element transcriptional activity.
Pin1 is a unique regulator, which catalyzes the conversion of a specific phospho-Ser/Thr-Pro-containing motif in target proteins. Herein, we identified CRTC2 as a Pin1-binding protein by overexpressing Pin1 with Myc and FLAG tags in mouse livers and subsequent purification of the complex containing Pin1. The association between Pin1 and CRTC2 was observed not only in overexpression experiments but also endogenously in the mouse liver. Interestingly, Ser(136) in the nuclear localization signal of CRTC2 was shown to be involved in the association with Pin1. Pin1 overexpression in HepG2 cells attenuated forskolin-induced nuclear localization of CRTC2 and cAMP-responsive element (CRE) transcriptional activity, whereas gene knockdown of Pin1 by siRNA enhanced both. Pin1 also associated with CRTC1, leading to their cytosol localization, essentially similar to the action of CRTC2. Furthermore, it was shown that CRTC2 associated with Pin1 did not bind to CREB. Taken together, these observations indicate the association of Pin1 with CRTC2 to decrease the nuclear CBP·CRTC·CREB complex. Indeed, adenoviral gene transfer of Pin1 into diabetic mice improved hyperglycemia in conjunction with normalizing phosphoenolpyruvate carboxykinase mRNA expression levels, which is regulated by CRE transcriptional activity. In conclusion, Pin1 regulates CRE transcriptional activity, by associating with CRTC1 or CRTC2.
Nakatsu
Yusuke
Y
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Sakoda
Hideyuki
H
Departments of Internal Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Kushiyama
Akifumi
A
Institute for Adult Disease, Asahi Life Foundation, Tokyo 100-0006, Japan.
Ono
Hiraku
H
Departments of Internal Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Fujishiro
Midori
M
Departments of Internal Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Horike
Nanao
N
Physiological Chemistry and Metabolism, Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Yoneda
Masayasu
M
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Ohno
Haruya
H
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Tsuchiya
Yoshihiro
Y
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Kamata
Hideaki
H
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Tahara
Hidetoshi
H
Department of Cellular and Molecular Biology, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima 734-8553, Japan.
Isobe
Toshiaki
T
Center for Priority Areas, Tokyo Metropolitan University, Hachioji, Tokyo 192-0397, Japan.
Nishimura
Fusanori
F
Department of Dental Science for Health Promotion, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima 734-8553, Japan.
Katagiri
Hideki
H
Division of Molecular Metabolism and Diabetes, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
Oka
Yoshitomo
Y
Division of Molecular Metabolism and Diabetes, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.
Fukushima
Toshiaki
T
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan.
Takahashi
Shin-Ichiro
SI
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Kurihara
Hiroki
H
Physiological Chemistry and Metabolism, Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Uchida
Takafumi
T
Department of Molecular Cell Biology, Graduate School of Agricultural Science, Tohoku University, Sendai, Miyagi 981-8555, Japan.
Asano
Tomoichiro
T
From the Department of Medical Science, Graduate School of Medicine, University of Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima City, Hiroshima 734-8553, Japan. Electronic address: asano-tky@umin.ac.jp.
D021581:Active Transport, Cell Nucleus / Q000187:drug effects / Q000502:physiology; D000818:Animals; D050882:CREB-Binding Protein / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D055785:Gene Knockdown Techniques; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver / Q000378:metabolism; D051379:Mice; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism*; D019696:Peptidylprolyl Isomerase / Q000235:genetics / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology*
C115163:CREB1 protein, human; C500984:CRTC1 protein, human; C493245:CRTC2 protein, human; C492004:Creb1 protein, mouse; C504923:Crtc2 protein, mouse; D017362:Cyclic AMP Response Element-Binding Protein; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019913:Nuclear Localization Signals; D015534:Trans-Activators; D014157:Transcription Factors; D005576:Colforsin; D000242:Cyclic AMP; D050882:CREB-Binding Protein; C494363:CREBBP protein, human; C494364:Crebbp protein, mouse; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase; C000606850:Pin1 protein, mouse