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Molecular Cloning, Chromosomal Mapping, and Characterization of the Human Cardiac-Specific Homeobox Gene hCsx

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Abstract

Background

Csx/Nkx2.5, a murine nonclustered homeobox gene expressed primarily in the heart, has significant sequence similarity to the Drosophila tinman gene. Tinman is essential for heart and gut formation in Drosophila. Targeted mutation in the mouse gene, Csx/Nkx25, arrests cardiac development during early embryonic stages, suggesting an evolutionary conservation in cardiogenesis.

Materials and Methods

We have isolated and characterized a human homolog, hCsx, from an adult cardiac cDNA library. Northern blotting and ribonuclease protection was used to define the pattern of expression during normal development and in disease states. Chromosomal localization of the gene was determined by somatic cell hybrid analysis and fluorescent in situ hybridization.

Results

The predicted amino acid sequence of hCsx has 87% overall homology to the murine gene with 100% identity in the homeodomain. The homeodomain sequence of hCsx is 95% identical to its Xenopus homolog, and 65% to tinman. hCsx mRNA was detected exclusively in the heart. hCsx transcript was detected at 12 weeks in human embryonic heart, the earliest time point examined, and was up-regulated 5-fold between 12 and 19 weeks. There was no significant alteration of hCsx message level in the myocardium of 14 patients with end stage heart failure compared to a normal control. The human gene mapped to the distal portion of chromosome 5, the 5q34–q35 region. This defines a new synteny region between human chromosome 5q and the t-locus of mouse chromosome 17, where the mouse Csx gene is located.

Conclusions

hCsx, the human homolog of Drosophila tinman, is expressed in heart in a tissue restricted manner. Distal 5q trisomies produce several phenotypic abnormalities, including a high incidence of congenital heart disease. Isolation of the hCsx gene will allow further studies of mutations in this gene and their potential associations with some forms of congenital heart disease in humans.

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Acknowledgments

We thank Ann Burgess, Madhu Prasad, and Mary Richmond for excellent technical assistance; T. Takahashi and P. D. Allen for RNA samples; and N. G. Copeland, T. W. Glover, S. A. Camper, and D. Law for helpful comments on the manuscript. This work was supported in part by grants from the National Institutes of Health and the American Heart Association to SI.

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Author notes

  1. David Turbay

    Present address: Division of Immunogenetics, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA, 02115, USA

Authors and Affiliations

  1. Cardiovascular Research Center and Department of Internal Medicine, The University of Michigan Medical Center, Ann Arbor, Michigan, USA

    David Turbay, Kristina McQuate Blanchard & Seigo Izumo

  2. Department of Pediatrics and Communicable Diseases, The University of Michigan Medical Center, Ann Arbor, Michigan, USA

    Stephanie Burns Wechsler

  3. Department of Biological Chemistry, 7220 MSRB III, The University of Michigan Medical Center, 1150 West Medical Center Drive, Ann Arbor, Michigan, 48109-0644, USA

    Seigo Izumo

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  1. David Turbay
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Contributed by T. Yamada on October 11, 1995.

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Turbay, D., Wechsler, S.B., Blanchard, K.M. et al. Molecular Cloning, Chromosomal Mapping, and Characterization of the Human Cardiac-Specific Homeobox Gene hCsx. Mol Med 2, 86–96 (1996). https://doi.org/10.1007/BF03402205

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