ABSTRACT
We have isolated a gene from a cDNA library generated from the thymus of a mouse with severe combined immune deficiency, termed FKBP9, that encodes a protein related to FK506-binding protein 6 (65 kDa, FKBP65). FKBP9 contains four peptidyl-prolyl cis-trans isomerase (PPIase) signature and two EF-hand domains which is identical to FKBP6/65 in overall structural organization. However, the two proteins share only 66% amino acid identity. FKBP9 is expressed at high levels in mouse heart, muscle, lung, and kidney. While FKBP6 was previously mapped to chromosome 11, the Fkbp9 gene was mapped to mouse chromosome 6 by analysis of a multilocus cross. These results identify a new member of the mouse FKBP protein family located on a separate chromosome.
Subject(s)
Peptidylprolyl Isomerase , Tacrolimus Binding Proteins/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Gene Expression , Mice , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Sequence Homology, Amino Acid , Tissue DistributionABSTRACT
We cloned and sequenced a mouse gene encoding a new type of membrane bound serine protease (epithin) containing a multidomain structure. The initial cDNA clone was found previously in a polymerase chain reaction (PCR)-based subtractive library generated from fetal thymic stromal cells, and the message was shown to be highly expressed in a thymic epithelial nurse cell line. A clone isolated from a severe combined immunodeficiency (SCID) thymus library and extended to its full length at the 5' end with the RACE technique contains an open reading frame of 902 amino acids. Based on the sequence of this clone, the predicted protein structure is a type II membrane protein with a C-terminal serine protease domain linked to the membrane by four low density lipoprotein receptor modules and two CUB domains. High message expression by northern blotting was detected in intestine, kidney, lung, SCID, and Rag-2(-/-) thymus, and 2-deoxyguanosine-treated fetal thymic rudiment, but not in skeletal muscle, liver, heart, testis, and brain. Sorted MHC class II+ and II- fetal thymic stromal cells were positive for expression by reverse transcriptase-PCR, whereas CD45(+) thymocytes were not. The gene was found in chicken and multiple mammalian species under low stringency Southern hybridization conditions. Under high stringency conditions, only a single gene per haploid genome was identified in the mouse. This gene, Prss14 (protease, serine, 14), was mapped to mouse chromosome 9 and is closely linked to the Fli1 (Friend leukemia integration 1) gene.
Subject(s)
Serine Endopeptidases/genetics , Thymus Gland/enzymology , Amino Acid Sequence , Animals , Chromosome Mapping , Cloning, Molecular , DNA, Complementary/genetics , Membrane Proteins , Mice , Mice, SCID , Mice, Transgenic , Molecular Sequence Data , Protein Conformation , Receptors, LDL/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Stromal Cells , Thymus Gland/cytology , Tissue DistributionABSTRACT
The Asian mouse Mus castaneus is resistant to infection by the polytropic mink cell focus-inducing (MCF) subgroup of murine leukemia viruses (MuLVs). Genetic crosses showed this recessive resistance to be governed by a single gene that maps at or near the gene encoding the polytropic viral receptor, Rmc1. To investigate this resistance, we mated M. castaneus with mice carrying the wild mouse Sxv variant of the Rmc1 receptor that allows infection by xenotropic as well as polytropic virus. Unlike other F1 hybrids of M. castaneus, these F1 mice were resistant to both xenotropic and polytropic classes of MuLVs. Analysis of backcrossed progeny of the F1 hybrids mated to Sxv mice indicates that resistance is due to inheritance of two M. castaneus genes. Cells from individual backcross mice were also examined for cell surface antigen by fluorescence-activated cell sorter analysis with monoclonal antibodies reactive with xenotropic or MCF virus env glycoproteins. A correlation was observed between virus resistance and antigen, suggesting that virus resistance is due to expression of endogenous viral envelope genes that interfere with infection by exogenous virus. Since the inbred strain Rmc1 receptor remains functional in the presence of these M. castaneus genes, and since M. castaneus contains multiple copies of xenotropic MuLV env genes, we suggest that these resistance genes control expression of xenotropic env glycoprotein that interferes with exogenous virus in cells containing the Sxv variant of Rmc1.
Subject(s)
Membrane Proteins , Mink Cell Focus-Inducing Viruses/immunology , Muridae/immunology , Receptors, Virus/metabolism , Animals , Cells, Cultured , Female , Immunity, Innate , Male , Mice , Mice, Inbred BALB C , Mice, Inbred DBA , Mink Cell Focus-Inducing Viruses/metabolism , Muridae/genetics , Receptors, G-Protein-Coupled , Receptors, Virus/genetics , Viral Envelope Proteins/immunology , Xenotropic and Polytropic Retrovirus ReceptorABSTRACT
We describe here a novel member of the slow-kinetics immediate-early gene family. Ier5 is an intronless gene, encoding a serum- and growth factor-inducible message of 2123 nucleotides that is present in a wide variety of tissues. The predicted open reading frame encodes a 308-amino-acid, highly proline-rich protein with homology to the amino terminus of the immediate-early gene pip92/Ier2/ETR101. Ier5 is predicted to be a nuclear protein and contains a PEST-like sequence, suggesting rapid protein degradation. Multiple phosphorylation sites are present. Ier5 shows growth factor induction kinetics similar to that of pip92/Ier2/ETR101, but unlike pip92/Ier2/ETR101 does not appear to require phosphokinase C activity for transcriptional activation. The sequence of the promoter region of Ier5 was determined and examined for transcription factor binding sites thought to mediate serum and growth factor response. Multiple AP-1 sites and an Ets-1 site were observed, but the CArG and CArG-like boxes of the serum response element were absent. The predicted nuclear localization of Ier5, coupled with the potential for rapid regulation by phosphorylation and/or degradation, suggests that Ier5 may play an important role in mediating the cellular response to mitogenic signals.
Subject(s)
Genes, Immediate-Early/genetics , Immediate-Early Proteins/genetics , Nuclear Proteins , 3T3 Cells , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Brain Chemistry/genetics , Dose-Response Relationship, Drug , Growth Substances/genetics , Growth Substances/pharmacology , Mice , Molecular Sequence Data , Sequence Analysis, DNA , Time Factors , Transcription FactorsABSTRACT
Chemokines refer to a rapidly expanding family of small cytokines whose primary function is recruitment of leukocytes to inflammatory sites. These are known to bind to seven-transmembrane-domain containing receptors. A cDNA clone, CHEMR1, resembling the typical G protein-coupled receptor, was isolated from a mouse cytotoxic T-lymphocyte (CTL) library. Northern blot analysis in mouse cell lines suggests that its expression is found in a variety of cells, including T cells, B cells, and macrophages. The CHEMR1 gene Scya3r2 is a single-copy gene whose open reading frame may be in a single exon and maps to the distal region of mouse Chr 9 where the mouse macrophage inflammatory protein-1alpha (MIP-1alpha) receptor gene Scya3r and two related C-C chemokine receptor-like genes reside. Amino acid sequence comparison shows that CHEMR1 is 84% identical to human CCR-4, indicating that CHEMR1 is likely to be a mouse CCR-4. Binding assays using 125I-labeled C-C chemokines in mammalian cells indicated that CHEMR1 did not bind MIP-1alpha, RANTES, or MIP-1beta, whereas CCR-1 binds MIP-1alpha and RANTES. Our result is different from the reported properties of human CCR-4. This suggests that CHEMR1 may be a receptor for unidentified C-C chemokine or a low-affinity receptor for MIP-1alpha.
Subject(s)
Chemokines, CC , Chemokines/metabolism , DNA, Complementary/genetics , Fungal Proteins/genetics , Genes , Mice/genetics , Receptors, Chemokine , Ribonucleases , Saccharomyces cerevisiae Proteins , T-Lymphocytes, Cytotoxic/chemistry , Transcription Factors/genetics , Amino Acid Sequence , Animals , Base Sequence , Chemokine CCL17 , Chemokine CCL3 , Chemokine CCL4 , Chemokine CCL5/metabolism , Chromosome Mapping , Cloning, Molecular , Fungal Proteins/metabolism , Gene Library , Humans , Macrophage Inflammatory Proteins/metabolism , Molecular Sequence Data , Organ Specificity , Receptors, CCR1 , Receptors, Cytokine/metabolism , Recombinant Fusion Proteins/metabolism , Sequence Alignment , Sequence Homology, Amino Acid , Species Specificity , Substrate Specificity , Transcription Factors/metabolismABSTRACT
Fv4 is an endogenous defective murine leukaemia virus (MuLV) which expresses high levels of an envelope protein (Env) closely related to that of the ecotropic class of MuLVs. Mice bearing the natural Fv4 gene or a transgenic version are resistant to infection by ecotropic MuLVs. Fv4 mice secrete the surface peptide (SU) of the Fv4 Env in their serum and this secreted Env can block infection of NIH3T3 cells. To study the secretion of Fv4, we metabolically labelled cells expressing Fv4 Env or Env from infectious MuLVs and followed synthesis, glycosylation, proteolytic processing and secretion of Env species. We found no difference in the kinetics of synthesis or processing of Fv4 Env compared to the envelopes of infectious MuLVs, but Fv4 Env associated more weakly with its transmembrane anchor and was shed from the surface of cells.
Subject(s)
Immunity, Innate/genetics , Leukemia Virus, Murine/metabolism , Leukemia, Experimental/metabolism , Membrane Proteins/metabolism , Retroviridae Infections/metabolism , Tumor Virus Infections/metabolism , Animals , Immunity, Innate/immunology , Leukemia, Experimental/immunology , Membrane Proteins/genetics , Membrane Proteins/immunology , Mice , Retroviridae Infections/immunology , Tumor Virus Infections/immunologyABSTRACT
We have cloned and characterized the organization of the mouse orphan nuclear receptor Nurr1 gene. The Nurr1 gene is approximately 7 kb long, contains eight exons and seven introns, and mapped to mouse chromosome 2. Although the exon/intron structure of Nurr1 is nearly identical to that of Nur77, Nurr1 possesses an additional untranslated exon. Primer extension was used to identify two major transcription initiation sites mapped 37 nucleotides apart in the first untranslated exon. Functional studies of chimeric Nurr1-luciferase reporter genes delineated the promoter region and underscored the importance of the +1 transcription start site. Sequence analysis of the 5' flanking region surrounding +1 revealed several possible response elements such as a hexanucleotide glucocorticoid binding site, a cAMP-response element, a CArG box, and two c-Jun-binding sites. These data help to explain the different response characteristics of two closely related early response genes, Nurr1 and Nur77.
Subject(s)
Chromosome Mapping , DNA-Binding Proteins , Promoter Regions, Genetic , Receptors, Cytoplasmic and Nuclear/genetics , Transcription Factors/genetics , Animals , Base Sequence , Binding Sites , DNA, Complementary , Mice , Molecular Sequence Data , Nuclear Receptor Subfamily 4, Group A, Member 2 , Peptide Chain Initiation, Translational , Transcription, GeneticABSTRACT
A cell line designated CUMO-2 has been established from an undifferentiated ovarian carcinoma. The s.c. injection of cells into nude mice gave rise to fast-growing tumors, while the i.p. route induced a peritoneal carcinomatosis with ascites. Histopathologically, the transplanted s.c. tumors closely resembled the original tumor, but tumors developed in the peritoneal cavity were highly anaplastic. The epithelial nature of the cells was confirmed by ultrastructural analysis. Sequential cytogenetic analyses on early and late passages revealed highly aneuploid tumor cells with consistent structural aberrations of chromosomes 1, 3, 8 and 11. CUMO-2 cells were found to produce CA 125 in vitro and in vivo. Cytosol estrogen receptor (ER) was found but progesterone receptor (PR) was not measured. HLA typing indicated the presence of DR8 and DQw4. A gonadotropin-releasing hormone (Gn-RH) analog inhibited cell growth and Gn-RH receptor mRNA was detected by reverse transcription/polymerase chain reaction in this cell line. Administration of transforming growth factor beta 1 inhibited both cell growth and c-myc mRNA expression. This cell line demonstrated a conformational band shift in exon 7 of the p53 gene. It was a frameshift mutation.
Subject(s)
Carcinoma/pathology , Cell Line , Ovarian Neoplasms/pathology , Animals , CA-125 Antigen/biosynthesis , Carcinoma/chemistry , Carcinoma/genetics , Cell Differentiation , Cell Division , Chromosome Banding , Female , Gene Expression , Genes, myc , Growth Inhibitors/pharmacology , HLA Antigens/analysis , Humans , Mice , Mice, Nude , Middle Aged , Neoplasm Metastasis , Neoplasm Transplantation , Ovarian Neoplasms/chemistry , Ovarian Neoplasms/genetics , Polymorphism, Single-Stranded Conformational , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Receptors, Estrogen/analysis , Receptors, LHRH/genetics , Transforming Growth Factor beta/pharmacologyABSTRACT
Using a previously isolated mouse centromere protein A (Cenpa) probe, we have localized the gene to the proximal region of mouse Chromosome 5, between the known Il6 and Yes1 loci near [Adra2C-D5H4S43-Hdh]. Comparison of this localization with that of human CENPA, which maps to chromosome 2, is consistent with the presence of a new region of conserved synteny between the two species.
Subject(s)
Autoantigens , Chromosomal Proteins, Non-Histone/genetics , Mice/genetics , Animals , Centromere Protein A , Chromosome Mapping , Chromosomes, Human, Pair 5 , HumansABSTRACT
To determine chromosome positions for 10 mouse phospholipase C (PLC) genes, we typed the progeny of two sets of genetic crosses for inheritance of restriction enzyme polymorphisms of each PLC. Four mouse chromosomes, Chr 1, 11, 12, and 19, contained single PLC genes. Four PLC loci, Plcb1, Plcb2, Plcb4, and Plcg1, mapped to three sites on distal mouse Chr 2. Two PLC genes, Plcd1 and Plcg2, mapped to distinct sites on Chr 8. We mapped the human homologs of eight of these genes to six chromosomes by analysis of human x rodent somatic cell hybrids. The map locations of seven of these genes were consistent with previously defined regions of conserved synteny; Plcd1 defines a new region of homology between human Chr 3 and mouse Chr 8.
Subject(s)
Chromosome Mapping , Chromosomes, Human, Pair 3 , Polymorphism, Restriction Fragment Length , Type C Phospholipases/genetics , Animals , Base Sequence , Cricetinae , Crosses, Genetic , DNA Primers , DNA Restriction Enzymes , Genetic Markers , Humans , Hybrid Cells , Mice , Molecular Sequence Data , Polymerase Chain ReactionSubject(s)
Microtubule-Associated Proteins , Proteins/physiology , Tyrosine 3-Monooxygenase , 1-Alkyl-2-acetylglycerophosphocholine Esterase , 14-3-3 Proteins , Animals , Base Sequence , Brain/abnormalities , DNA Probes/genetics , DNA, Complementary , Humans , Mice , Molecular Sequence Data , Proteins/genetics , Rats , Sequence Homology , SyndromeABSTRACT
Transgenic mice bearing a murine retroviral envelope transgene (Fv4) have Fv4 gp70env (SU) in their serum in amounts sufficient to block infection by ecotropic virus in vitro. Fv4 Env in serum is derived largely but not exclusively from hematopoietic cells. Tail cells from Fv4 mice and cell lines transduced with the Fv4 env transgene synthesize both components of the envelope protein (gp70 SU and p15E TM) but secrete the gp70 moiety, in the absence of retroviral particles. Blocking of the ecotropic viral receptor by secreted gp70 SU may contribute to resistance to retroviral infection in these mice.
Subject(s)
Friend murine leukemia virus/physiology , Membrane Glycoproteins , Moloney murine leukemia virus/physiology , Receptors, Virus , Retroviridae Proteins, Oncogenic/blood , Viral Envelope Proteins/blood , Viral Interference , 3T3 Cells , Animals , Antibodies, Viral/immunology , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/metabolism , Cell Line , Female , Friend murine leukemia virus/genetics , HeLa Cells , Hematopoietic Stem Cells/metabolism , Humans , Male , Membrane Proteins/antagonists & inhibitors , Membrane Proteins/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Moloney murine leukemia virus/genetics , Retroviridae Proteins, Oncogenic/genetics , Retroviridae Proteins, Oncogenic/immunology , Retroviridae Proteins, Oncogenic/metabolism , Viral Envelope Proteins/genetics , Viral Envelope Proteins/immunology , Viral Envelope Proteins/metabolismSubject(s)
Chromosome Mapping , Sialoglycoproteins/genetics , Animals , Base Sequence , Bone and Bones/chemistry , Cell Line , Cricetinae , DNA Primers , Female , Humans , Male , Mice , Molecular Sequence Data , OsteopontinABSTRACT
Cultured cells derived from the wild mouse species Mus castaneus were found to be uniquely resistant to exogenous infection by polytropic mink cell focus-forming (MCF) murine leukemia viruses (MuLVs). This MCF MuLV resistance is inherited as a genetically recessive trait in the progeny of F1 crosses between M. castaneus and MCF MuLV-susceptible laboratory mice. Examination of the progeny of backcrosses demonstrated that susceptibility is inherited as a single gene which maps to chromosome 1. The map location of this gene places it at or near the locus Rmc1, the gene encoding the receptor for MCF/xenotropic MuLVs, suggesting that resistance is mediated by the M. castaneus allele of this receptor.
Subject(s)
Mink Cell Focus-Inducing Viruses/physiology , Muridae/genetics , Animals , Animals, Wild , Cell Line , Cells, Cultured , Chromosome Mapping , Genes, Recessive , Immunity, Innate/genetics , Mice , Mice, Inbred AKR , Mice, Inbred DBA , Mink Cell Focus-Inducing Viruses/genetics , Muridae/virology , Receptors, Virus/genetics , Xenotropic and Polytropic Retrovirus ReceptorABSTRACT
We have isolated genomic and cDNA clones of Brca1, a mouse homolog of the recently cloned breast cancer-associated gene, BRCA1. Brca1 encodes an 1812-amino-acid protein with a conserved zinc finger domain and significant homology to the human protein. Brca1 maps to Chromosome 11 within a region of conserved synteny with human chromosome 17, consistent with the mapping of the human gene to 17q21. Brca1 transcripts are expressed in a variety of cultured cells but reveal a specific and dynamic expression pattern during embryonic development. For example, expression is observed first in the otic vesicle of embryonic day 9.5 (E9.5) embryos. This expression diminishes and is replaced by expression in the neuroectoderm at E10.5. By E11-12.5, higher levels are observed in differentiating keratinocytes and in whisker pad primordia. Transcripts also become evident in epithelial cells of the E14-17 kidney. Brca1 expression occurs in differentiating epithelial cells of several adult organs as well, suggesting a general role in the functional maturation of these tissues. Consistent with this, Brca1 transcripts are expressed in both alveolar and ductal epithelial cells of the mammary gland. During pregnancy, there is a large increase in Brca1 mRNA in mammary epithelial cells, an increase that parallels their functional differentiation. Because high rates of breast cancer are associated with loss of BRCA1 in humans, it is possible that this gene provides an important growth regulatory function in mammary epithelial cells. In addition, increased transcription of mammary Brca1 during pregnancy might contribute, in part, to the reduced cancer risk associated with exposure to pregnancy and lactation.