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Volume 271, Number 24, Issue of June 14, 1996 pp. 14567-14571
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Molecular Cloning of a Novel Thyroid Hormone-responsive Gene, ZAKI-4, in Human Skin Fibroblasts

(Received for publication, August 22, 1995, and in revised form, March 19, 1996)

Takashi Miyazaki Dagger § , Yasuhiko Kanou , Yoshiharu Murata Dagger , Sachiko Ohmori Dagger , Toshimitsu Niwa § , Kenji Maeda § , Hideki Yamamura and Hisao Seo Dagger

From the Dagger  Department of Endocrinology and Metabolism and the  Department of Teratology and Genetics, Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-01, Japan and the § Department of Internal Medicine, Nagoya University Branch Hospital, Nagoya 461, Japan

Utilizing a method called ``differential display of mRNAs by means of polymerase chain reaction'', the cDNA fragment of a thyroid hormone-responsive gene ZAKI-4 was cloned from cultured human skin fibroblasts. Northern blot analysis revealed that there were two ZAKI-4 mRNA species (3.4 and 1.4 kilobases (kb)), and they were up-regulated by a physiological concentration of triiodothyronine (T3). This T3 effect was abolished by the treatment with cycloheximide, indicating the possibility that gene ZAKI-4 is regulated by T3 in an indirect fashion, through an intermediate product of T3, rather directly by T3 itself. No effect of T3 on ZAKI-4 mRNA stability suggested that T3 induces the mRNA at the transcriptional level. Rapid amplification of cDNA ends confirmed the presence of two mRNA species. ZAKI-4 mRNA was detected in heart, brain, liver, and skeletal muscle but not in placenta, lung, kidney and pancreas. In skin fibroblasts and skeletal muscle, 3.4-kb mRNA was the major species, whereas 1.4-kb mRNA was dominant in heart, brain, and liver. The sequence analysis suggested that the two mRNA species arise from alternative polyadenylation and code a single protein of 192 amino acids. No homologous protein sequence was found in a data base. Elucidation of the function of ZAKI-4 gene product will provide new insights into an important role of T3 in various organs.


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