Molecular cloning, expression and characterization of the zebrafish bram1 gene, a BMP receptor-associated molecule.
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 approximately approximately 98% homology with human and mouse BRAM1, and approximately approximately 64% homology with C. elegans BRA-1 and BRA-2. The bram1 gene, composed of five exons and four introns, spans approximately approximately 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.
Wu
Kang-mai
KM
Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan.
Huang
Chang-jen
CJ
Hwang
Sheng-ping L
SP
Chang
Yu-sun
YS
D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D019556:COS Cells; D017173:Caenorhabditis elegans; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D020224:Expressed Sequence Tags; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000737:chemistry* / Q000235:genetics*
D048868:Adaptor Proteins, Signal Transducing; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D029961:Zebrafish Proteins; C512736:zmynd11 protein, zebrafish