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Published 2 February 2004. doi:10.1083/jcb.200309159
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 164, Number 3, 395-405
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Article

 A novel LIM protein Cal promotes cardiac differentiation by association with CSX/NKX2-5

Hiroshi Akazawa1, Sumiyo Kudoh2, Naoki Mochizuki3, Noboru Takekoshi2, Hiroyuki Takano1, Toshio Nagai1, and Issei Komuro1

1 Department of Cardiovascular Science and Medicine, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan
2 Department of Cardiology, Kanazawa Medical University, Ishikawa 920-0265, Japan
3 Department of Structural Analysis, National Cardiovascular Center Research Institute, Osaka 565-8565, Japan

Address correspondence to Issei Komuro, Dept. of Cardiovascular Science and Medicine, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan. Tel.: 81-43-226-2097. Fax: 81-43-226-2557. email: komuro-tky{at}umin.ac.jp

The cardiac homeobox transcription factor CSX/NKX2-5 plays an important role in vertebrate heart development. Using a yeast two-hybrid screening, we identified a novel LIM domain–containing protein, named CSX-associated LIM protein (Cal), that interacts with CSX/NKX2-5. CSX/NKX2-5 and Cal associate with each other both in vivo and in vitro, and the LIM domains of Cal and the homeodomain of CSX/NKX2-5 were necessary for mutual binding. Cal itself possessed the transcription-promoting activity, and cotransfection of Cal enhanced CSX/NKX2-5–induced activation of atrial natriuretic peptide gene promoter. Cal contained a functional nuclear export signal and shuttled from the cytoplasm into the nucleus in response to calcium. Accumulation of Cal in the nucleus of P19CL6 cells promoted myocardial cell differentiation accompanied by increased expression levels of the target genes of CSX/NKX2-5. These results suggest that a novel LIM protein Cal induces cardiomyocyte differentiation through its dynamic intracellular shuttling and association with CSX/NKX2-5.

Key Words: cardiogenesis; homeobox transcription factor; LIM domain; nucleocytoplasmic transport; transcriptional regulation

Abbreviations used in this paper: ANP, atrial natriuretic peptide; Ca2+, calcium; Cal, CSX-associated LIM protein; CRP, cysteine-rich protein; LMB, leptomycin B; LPP, lipoma preferred partner; NES, nuclear export signal; SERCA2, sarcoplasmic reticulum Ca2+-ATPase 2; trip6, thyroid receptor interacting protein 6.


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