TY - JOUR
T1 - Identification, chromosomal assignment, and expression analysis of the human homeodomain-containing gene Orthopedia (OTP)
AU - Lin, Xu
AU - State, Matthew W.
AU - Vaccarino, Flora M.
AU - Greally, John
AU - Hass, Melanie
AU - Leckman, James F.
N1 - Funding Information:
This work was supported in part by the Korzac Foundation and a grant from the National Institute of Mental Health (MH49351). The authors thank M. Ding for her help in the characterization of the OTP antiserum and A. Simeone for providing the murine Otp probe as well as the Otp expression vectors. We extend our appreciation to D. Ward, P. Burbach, M. Biggin, E. Burnett, M. Murtha, and P. J. Lombroso for their comments during the preparation of the manuscript. We also thank Drs. A Swaroop, D. P. Bick, R. Robins, K. Kagan-Hallet, and J. McGill for their roles in collecting and preparing the fetal brain library used in this study. C. Howe assisted in the collection and preparation of the human fetal tissue used in the expression study.
PY - 1999/8/15
Y1 - 1999/8/15
N2 - Homeodomain (HD) genes are helix-turn-helix transcription factors that play key roles in the specification of cell fates. In the central nervous system (CNS), HD genes not only position cells along an axis, but also specify cell migration patterns and may influence axonal connectivity. In an effort to identify novel HD genes involved in the development of the human CNS, we have cloned, characterized, and mapped the human homologue of the murine HD gene Orthopedia (Otp), whose product is found in multiple cell groups within the mouse hypothalamus, amygdala, and brain stem. Human cDNA and genomic libraries were screened with probes derived from mouse Otp sequences to find the human homologue, OTP. The deduced amino acid sequence of the open reading frame of the human cDNA is 99% homologous to mouse Otp and demonstrates a high degree of conservation when compared to sea urchin and Drosophila. OTP was mapped to human chromosome 5q13.3 using radiation hybrid panel mapping and fluorescence in situ hybridization. Flanking markers were identified from YAC clones containing OTP. A single putative OTP gene product was found in 17-week human fetal brain tissue by Western blot analysis using a novel polyclonal antibody raised against a conserved 13- amino-acid sequence at the C-terminus of the OTP protein. Expression in the developing human hypothalamus was confirmed by immunohistochemistry.
AB - Homeodomain (HD) genes are helix-turn-helix transcription factors that play key roles in the specification of cell fates. In the central nervous system (CNS), HD genes not only position cells along an axis, but also specify cell migration patterns and may influence axonal connectivity. In an effort to identify novel HD genes involved in the development of the human CNS, we have cloned, characterized, and mapped the human homologue of the murine HD gene Orthopedia (Otp), whose product is found in multiple cell groups within the mouse hypothalamus, amygdala, and brain stem. Human cDNA and genomic libraries were screened with probes derived from mouse Otp sequences to find the human homologue, OTP. The deduced amino acid sequence of the open reading frame of the human cDNA is 99% homologous to mouse Otp and demonstrates a high degree of conservation when compared to sea urchin and Drosophila. OTP was mapped to human chromosome 5q13.3 using radiation hybrid panel mapping and fluorescence in situ hybridization. Flanking markers were identified from YAC clones containing OTP. A single putative OTP gene product was found in 17-week human fetal brain tissue by Western blot analysis using a novel polyclonal antibody raised against a conserved 13- amino-acid sequence at the C-terminus of the OTP protein. Expression in the developing human hypothalamus was confirmed by immunohistochemistry.
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U2 - 10.1006/geno.1999.5882
DO - 10.1006/geno.1999.5882
M3 - Article
C2 - 10458915
AN - SCOPUS:0345624500
SN - 0888-7543
VL - 60
SP - 96
EP - 104
JO - Genomics
JF - Genomics
IS - 1
ER -