ABSTRACT
We have cloned the full length of a novel cDNA, named ring finger protein 21 (RNF21), composed of the RING finger-B box-coiled coil (RBCC) domain and the B30.2 domain, which are characteristic of the RBCC-B30.2 family. As a structural feature, the RNF21 cDNA possessed at least three kinds of isoforms, due to alternative splicing, consisting of the long form with the RBCC-RBCC-B30.2 domain, the medium form with the RBCC-B30.2 domain, and the short form with only the RBCC domain. Moreover, respective transcripts corresponding to the three isoforms were detected in various human organs by reverse transcription-PCR and Northern blot analyses. Interestingly, the medium form of the RNF21 mRNA expressed most predominantly was dramatically up-regulated within 8-16 h by interferon stimulation of HeLa cells. These findings suggest that RNF21 is a downstream gene that may mediate interferon's biological action.
Subject(s)
Carrier Proteins/genetics , Zinc Fingers/genetics , Alternative Splicing , Amino Acid Sequence , Animals , Blotting, Northern , COS Cells , Chromosome Banding , Chromosome Mapping , Chromosomes, Human, Pair 11/genetics , Cloning, Molecular , DNA, Complementary/chemistry , DNA, Complementary/genetics , Female , Gene Expression Regulation/drug effects , HeLa Cells , Humans , In Situ Hybridization, Fluorescence , Interferons/pharmacology , Male , Molecular Sequence Data , Protein Isoforms/genetics , Protein Structure, Tertiary , RNA, Messenger/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Repetitive Sequences, Amino Acid , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Tissue DistributionABSTRACT
We have previously identified an estrogen-responsive gene, efp (estrogen-responsive finger protein), by genomic binding-site cloning method. Here, we isolated a rat homologue of efp cDNA that encodes an open reading frame of 644 amino acids sharing high homology with human efp (69% identity at the protein level) and mouse efp (80% identity at the protein level). The efp protein has a RING finger, a variant type of zinc finger motif, B1 box and B2 box, each having a pair of zinc fingers, and coiled-coil domain, belonging to the RING finger-B box-Coiled Coil (RBCC) family. Several members of RBCC family including efp have characteristic C-terminal domain, forming a subfamily. Next, we detected efp mRNA in primary osteoblasts, one of estrogen target cells, derived from the calvariae of rat fetus. An anti-efp antibody revealed the efp protein is expressed and regulated by estrogen in the primary osteoblasts. Interestingly, the efp protein in primary osteoblasts is down-regulated by 1alpha,25-dihydroxyvitamin D(3) treatment that promotes the differentiation of the cells, whereas it is up-regulated by TGF-beta1 treatment that inhibits the differentiation of the cells. These findings suggest the possible involvement of the efp in the differentiation of osteoblastic cells.
Subject(s)
DNA-Binding Proteins/genetics , Osteoblasts/metabolism , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Calcitriol/pharmacology , Cell Differentiation/drug effects , Cell Differentiation/physiology , Cells, Cultured , Cloning, Molecular , DNA, Complementary/genetics , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/metabolism , Down-Regulation/drug effects , Estrogens/metabolism , Gene Expression Regulation , Humans , Mice , Molecular Sequence Data , Osteoblasts/cytology , Osteoblasts/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Sequence Homology, Amino Acid , Species Specificity , Transcription Factors/chemistry , Transcription Factors/metabolism , Transforming Growth Factor beta/pharmacology , Tripartite Motif Proteins , Ubiquitin-Protein Ligases , Zinc Fingers/geneticsABSTRACT
Although it is well known that estrogen exerts its effect in the brain, the direct target genes transcriptionally regulated by estrogen or rather estrogen receptor (ER) are almost unknown. During the search for estrogen receptor-binding sites from human CpG island library, we found one genomic DNA fragment corresponding to the putative 3'-untranslated region of human NMDA receptor subunit 2D (NR2D) gene. It contained at least four half palindromic estrogen responsive elements (hEREs) within two hundred nucleotides, which was conserved also in the rat. Interestingly, the NR2D mRNA is co-localized with ERalpha and/or ERbeta mRNA in a number of regions of rat brain. We have also demonstrated that NR2D mRNA is up-regulated in rat hypothalamus by estrogen possibly via hEREs identified here. Thus, we suggest that NR2D is one of the direct targets of estrogen receptors which are involved in reproductive as well as non-reproductive actions in the brain.
Subject(s)
Brain/physiology , Receptors, Estrogen/physiology , Receptors, N-Methyl-D-Aspartate/genetics , Animals , Base Sequence , Gene Targeting , Humans , Hypothalamus/physiology , Molecular Sequence Data , Rats , Rats, Sprague-Dawley , Sequence Homology, Nucleic AcidABSTRACT
Bfp (brain finger protein) is a member of the RING finger protein family, which is highly expressed in the brain. We have previously shown that one copy of the human bfp gene, mapped at 17p11.2, was actually deleted in six of six Smith-Magenis syndrome (SMS) patients. Now we have isolated the mouse bfp cDNA. Using in situ hybridization and immunohistochemistry, the distribution of mouse bfp mRNA and protein was identified especially in neural cells of the cerebral cortex, hippocampus, lateral amygdaloid nucleus, and ventromedial hypothalamus. In primary culture of the whole brain in a neonatal mouse, the Bfp protein was detected in both neuron and glial cells, and its subcellular localization was predominantly in the nucleus, but some amounts were also found in the cytoplasm. The bfp mRNA was also expressed strongly in the marginal zone of brain vesicles, optic stalk, and cartilage primordium, which are part of the critical tissues frequently involved in SMS patients, and in such tissues as nasal epithelium and primordium of follicles in a 13. 5-dpc embryo. Subsequently, its amount in the developing brain further increased during embryogenesis, reaching the highest level in the adult brain. These findings suggest a possibility that Bfp might be involved in the pathogenesis of Smith-Magenis syndrome as a regulator protein related to neural differentiation and function.
Subject(s)
Abnormalities, Multiple/genetics , Brain/metabolism , DNA-Binding Proteins/genetics , Intellectual Disability/genetics , Nerve Tissue Proteins/genetics , Zinc Fingers/genetics , Amino Acid Sequence , Animals , Blotting, Northern , Brain/embryology , Cells, Cultured , Chromosome Mapping , Cloning, Molecular , DNA, Complementary , DNA-Binding Proteins/analysis , DNA-Binding Proteins/chemistry , Fluorescent Antibody Technique , Gene Expression Regulation, Developmental , Humans , Immunohistochemistry , In Situ Hybridization , Mice , Molecular Sequence Data , Nerve Tissue Proteins/analysis , Nerve Tissue Proteins/chemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , SyndromeABSTRACT
RING finger is a variant zinc finger motif present in a new family of proteins including transcription regulators. Here, utilizing the polymerase chain reaction with degenerate primers, we isolated a genomic DNA fragment containing the RING finger motif. Using this fragment as a probe, we have identified a novel cDNA from rat testis library. Then, the human homologue of the terf cDNA was also isolated from a testis library. This gene was designated testis RING finger protein (terf) because the corresponding transcripts were detected almost exclusively in the testis by Northern blot analysis. Both cDNAs encode an open reading frame of 477 amino acids sharing high homology (74% identity at the protein level) between two species. The terf contains an N-terminal RING finger domain, one B-box domain, middle coiled-coil domain, and a C-terminal domain, belonging to the RING finger-B box-coiled coil (RBCC) family. Several RBCC proteins, such as PML, TIF1alpha and RFP, have transformation capabilities when found in chromosomal translocations. Among the members of the RBCC family, the terf shares highest homology (40% identity at the protein level) with RFP that is expressed only in the testis in normal tissues. Structural similarity raises the possibilities that the terf gene might be also involved in carcinogenesis or cell transformation.
Subject(s)
Carrier Proteins/genetics , Testis/metabolism , Transcription, Genetic , Zinc Fingers , Amino Acid Sequence , Animals , Base Sequence , Carrier Proteins/biosynthesis , Carrier Proteins/chemistry , Conserved Sequence , DNA, Complementary/chemistry , Gene Library , Humans , Male , Molecular Sequence Data , Open Reading Frames , Organ Specificity , RNA, Messenger/biosynthesis , Rats , Sequence Alignment , Sequence Homology, Amino Acid , Tripartite Motif Proteins , Ubiquitin-Protein LigasesABSTRACT
The bfp, a member of the RING finger family, has been shown to be predominantly expressed in brain and up-regulated in neural differentiation of P19 embryonic carcinoma cells. Chromosome mapping of the bfp gene by fluorescence in situ hybridization reveals that human BFP (ZNF179) is located at 17p11.2, mouse Bfp at 11B1.3, and rat BFP at 10q22. These results provide additional evidence that the mouse 11B region displays conserved linkage homology with the 17p11.2 region of the human genome and the 10q22 region of the rate genome.
Subject(s)
Brain/metabolism , Chromosome Mapping , Zinc Fingers/genetics , Amino Acid Sequence , Animals , Base Sequence , DNA, Complementary , Humans , In Situ Hybridization, Fluorescence , Mice , Molecular Sequence Data , Rats , Sequence Homology, Nucleic AcidABSTRACT
We have previously identified an estrogen-responsive gene, efp (estrogen-responsive finger protein), that encodes a putative zinc finger protein (Proc. Natl. Acad. Sci. USA 90: 11117-11121, 1993). The efp protein has a RING finger, a variant type of zinc finger motif, B1 box, and B2 box, each having a pair of zinc fingers, present in a family of apparent DNA-binding proteins. Some members of this family have transformation capabilities when found in chromosomal translocations. Chromosome mapping of the efp gene by fluorescence in situ hybridization reveals that human EFP (ZNF147) is located at 17q23.1 and that mouse Efp is located at 11C. These results provide additional evidence that the mouse 11C region displays conserved synteny with the 17q23.1 region of the human genome.
Subject(s)
Chromosomes, Human, Pair 17 , DNA-Binding Proteins/genetics , Genes , Mice/genetics , Transcription Factors/genetics , Animals , Breast Neoplasms/genetics , Chromosome Mapping , DNA, Complementary/genetics , Humans , In Situ Hybridization, Fluorescence , Multigene Family , Tripartite Motif Proteins , Ubiquitin-Protein Ligases , Zinc Fingers/geneticsABSTRACT
Estrogen receptor (ER)-binding fragments were isolated from human genomic DNA by using a recombinant ER protein. Using one of these fragments as a probe, we have identified an estrogen-responsive gene that encodes a putative zinc finger protein. It has a RING finger motif present in a family of apparent DNA-binding proteins and is designated estrogen-responsive finger protein (efp). efp cDNA contains a consensus estrogen-responsive element at the 3' untranslated region that can act as a downstream estrogen-dependent enhancer. Moreover, efp is regulated by estrogen as demonstrated at both the mRNA and the protein level in ER-positive cells derived from mammary gland. These data suggest that efp may represent an estrogen-responsive transcription factor that mediates phenotypic expression of the diverse estrogen action. Thus, the genomic binding-site cloning may be applicable for isolation of the target genes of other transcription factors.