Cloning and characterization of two nuclear receptors from the filarial nematode Brugia pahangi.
Nuclear receptors (NRs) encompass a superfamily of cytoplasmic/nuclear localized receptors that on ligand binding (or by phosphorylation) directly regulate the transcription of target genes. NRs are involved in many developmental processes, including moulting in insects and dauer larva formation in Caenorhabditis elegans. Here we report the isolation of two genes related to NRs from the filarial nematode Brugia pahangi. Bp-nhr-1 is a member of the NGF1-B subfamily of NRs and is expressed at very low levels in post-infective larval stage 3 (L3) after their transmission to the mammalian host. The second gene, Bp-nhr-2, is related to XR78E/F of Drosophila, a gene involved in the ecdysone response, over the region of its DNA-binding domain. cDNA and genomic clones have been isolated that correspond to Bp-nhr-2. The most striking feature of the encoded protein is that, although there is a DNA-binding domain similar to that of other NRs, the ligand-binding domain is absent. To investigate the pattern of transcription of Bp-nhr-2 in the filarial life cycle, semi-quantitative reverse-transcriptase-mediated PCR was performed; this analysis demonstrated that the gene is expressed in early stages after infection and in the adult and microfilariae, and is up-regulated just before the moult between L3 and L4 but is not expressed during the moult between L4 and adult. Antibodies raised against a peptide corresponding to the transactivation domain of Bp-nhr-2 demonstrate that the protein is expressed in microfilariae and adult samples and that another cross-reactive protein is expressed in these life-cycle stages.
Moore
J
J
Department of Veterinary Parasitology, Bearsden Road, University of Glasgow, Glasgow G61 1QH, U.K.
Devaney
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017179:Brugia pahangi / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017238:Genes, Helminth; D008969:Molecular Sequence Data; D018510:RNA, Helminth / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D029742:Caenorhabditis elegans Proteins; D017931:DNA Primers; D018509:DNA, Helminth; C105816:NHR-2 protein, C elegans; D018510:RNA, Helminth; D012333:RNA, Messenger; D018160:Receptors, Cytoplasmic and Nuclear