Molecular cloning, expression and characterization of the zebrafish bram1 gene, a BMP receptor-associated molecule |
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Authors: | Kang-mai Wu Chang-jen Huang Sheng-ping L Hwang Yu-sun Chang |
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Institution: | (1) Institute of Microbiology and Immunology, National Yang-Ming University, Shih-Pai 112, Taipei, Taiwan;(2) Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei, 11529, Taiwan;(3) Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, 11529, Taiwan;(4) Graduate Institute of Basic Medical Sciences, Chang Gung University, 259 Wen-Hwa 1st Road, Kwei-Shan, Taoyuan, 333, Taiwan |
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Abstract: | Summary We have identified a cDNA clone encoding BMP receptor-associated molecule 1 (BRAM1) from the zebrafish expressed sequence tag (EST) database. The 2606 bp full-length bram1 cDNA was cloned, and further confirmed by nucleotide sequencing. The zebrafish sequence encodes a protein of 195 amino acids with an evolutionarily conserved MYND domain, which displays ∼
∼98% homology with human and mouse BRAM1, and ∼
∼64% homology with C. elegans BRA-1 and BRA-2. The bram1 gene, composed of five exons and four introns, spans ∼
∼14 kb on linkage group 14 of the zebrafish genome. RT-PCR and whole mount in situ hybridization analyses disclosed that zebrafish BRAM1 is a maternal factor. The protein interacts directly with zebrafish BMP Receptor type IA, as observed from GST-pull down and co-immunoprecipitation assays. Furthermore, cotransfection of zebrafish BRAM1 with the corresponding BMP receptor resulted in down-regulation of BMP-mediated signaling. Our results collectively indicate that BRAM1 plays a biological role during zebrafish development. |
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Keywords: | BRAM1 BMPRIA whole-mount in situ hybridization real time RT-PCR zebrafish BMP signaling |
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