ABSTRACT
Three differentially expressed selectin genes (SELE, SELP, and SELL), important in the initial stages of leukocyte extravasation, have been reported in mammals. All three genes map close to the chemokine SCYC1 (small inducible cytokine subfamily C, member 1) in a large conserved chromosomal segment that extends from RXRG (retinoic acid receptor, gamma) to TNNT2 (troponin T2) on Chromosome (Chr) 1 in both human and mouse. In the mouse, we demonstrate that Sele is flanked by Prrx1 (paired-related homeobox gene 1) and Scyc1 and define the order of, and distances between, loci as centromere-Prrx1-(0.7+/-0.7 cM)-Sele-(1.2+/-0.9 cM)-Scyc1-telomere. In the chicken, we isolated BAC clones containing PRRX1, SELE, and SCYC1 and positioned them by fluorescent in situ hybridization. SELE and PRRX1 mapped to the short arm of chicken Chr 8 and SCYC1 mapped to the region equivalent to 1q11-1q13 on the long arm of chicken Chr 1. The location of SELE on chicken Chr 8 was independently established by linkage analysis of COM0185, an (AT)16 microsatellite locus identified in a BAC clone that contained SELE. COM0185 was linked to several loci that mapped to one end of chicken Chr 8, with the order of loci, and genetic distances (in cM) between them defined as MSU0435, MSU0325-(7.8+/-3.7)-COM0185-(5.8+/-3.2)-ROS0338-(9.6+/-4.0)-ABR0322-(3.8+/-2.6)-GLUL. We have therefore positioned an evolutionary breakpoint in mammals and chickens between SELE and SCYC1. Furthermore, comparative mapping analysis of the RXRG-TNNT2 chromosomal segment that is conserved on human and mouse Chr 1 indicates that it is divided into four segments in the chicken, each of which maps to a different chromosome.
Subject(s)
Chemokines, C , Chickens/genetics , Chromosomes , E-Selectin/genetics , Lymphokines/genetics , Sialoglycoproteins/genetics , Animals , Chromosome Mapping , Conserved Sequence , Evolution, Molecular , Expressed Sequence Tags , Genes , Genes, Homeobox , Genetic Linkage , Humans , In Situ Hybridization, Fluorescence , MiceABSTRACT
CD5 is a transmembrane glycoprotein on all T cells and on a subpopulation of B cells. Based on the analysis of chicken CD5-cDNA we have previously shown that the structure of the CD5 protein is conserved between species. Here we report the isolation and chromosomal mapping of the chicken CD5 gene. The gene spans 3.4 kb and is extremely compact with a high GC-nucleotide content. There are 10 exons and the introns are spliced out similarly to those in the human CD5 gene. Each of the three extracellular scavenger receptor cysteine-rich (SRCR) domains is encoded as an exon of its own, as is the proline-rich hinge region that separates the first two membrane-distal SRCR domains. The fluorescence in situ hybridization (FISH) technique was used to map the gene to chromosome five. This is the first report describing the organization of the CD5 gene from a nonmammalian species.
Subject(s)
CD5 Antigens/genetics , Chickens/genetics , Membrane Glycoproteins/genetics , Animals , Chickens/immunology , Chromosome MappingABSTRACT
Mottled mice have mutations in the copper-transporting ATPase Atp7a. They are proven models for the human disorder Menkes disease (MD), which results from mutations in a homologous gene. Mottled mice can be divided into three classes: class 1, in which affected males die before birth; class 2, in which affected males die in the early postnatal period; and class 3, in which affected males survive to adulthood. In humans, it has been shown that mutations that lead to a complete absence of functional protein cause classical MD, which is characterized by death of boys in early childhood. We hypothesized that the most severely affected mottled alleles would be the most likely to carry mutations equivalent to those causing classical MD and therefore undertook mutational analysis of several class 1 mottled alleles to assess whether these were appropriate models for the disease at the molecular level. Two novel mutations, a deletion of exons 11-14 in mottled spot and an insertion in exon 10 leading to missplicing in mottled candy, were identified. However, these are both "in-frame" mutations, as are the other eight Atp7a mutations reported to date, and therefore no frameshift or nonsense mutations have yet been associated with the mottled phenotype. This contrasts with the mutation spectrum associated with MD, emphasizing the need for caution when mottled mice are used as models for the clinical disorder.
Subject(s)
Adenosine Triphosphatases/genetics , Carrier Proteins/genetics , Cation Transport Proteins , Gene Deletion , Menkes Kinky Hair Syndrome/genetics , Recombinant Fusion Proteins , Alleles , Animals , Blotting, Southern , Copper-Transporting ATPases , Disease Models, Animal , Exons , Humans , Mice , Mice, Inbred C3H , Mice, Inbred C57BL , Models, Genetic , Mutation , Phenotype , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNASubject(s)
Carrier Proteins/genetics , Chromosomes, Artificial, Bacterial/genetics , Sex Chromosome Aberrations/genetics , Steroid Isomerases , Abnormalities, Multiple/genetics , Animals , Chondrodysplasia Punctata/genetics , Female , Humans , Male , Mice , Mice, Transgenic , Phenotype , X ChromosomeSubject(s)
Chickens/genetics , Conserved Sequence/genetics , Drosophila Proteins , Evolution, Molecular , Membrane Glycoproteins/genetics , Physical Chromosome Mapping , Receptors, Cell Surface/genetics , Animals , Crosses, Genetic , Female , Gene Order/genetics , Genetic Linkage/genetics , Humans , In Situ Hybridization, Fluorescence , Interleukin-8/genetics , Male , Mice , Toll-Like Receptor 2 , Toll-Like ReceptorsABSTRACT
A set of acute inflammation-regulated genes expressed in liver has been assigned to rat, mouse, and human chromosomes by detecting species-specific PCR amplicons in rat(x)mouse or mouse(x)hamster somatic cell hybrids or radiation hybrids or by in silico matches of corresponding rat cDNAs to various libraries of previously assigned rat, mouse, or human genes or expressed-sequence tags. This allowed us to assign 24, 22, and 21 inflammation-regulated genes to rat, mouse, and human chromosomes, respectively. From these assignments as well as those previously determined for a larger set of genes with an acute inflammation-regulated transcription in liver, we further investigated whether such genes are clustered onto given chromosomes. A cluster was found on rat Chromosome (Chr) 6q with a conserved synteny on mouse Chr 12 and human Chr 14q13-q32, and another cluster previously reported on human Chr 1q has been extended with five further genes. Our data suggest that during an acute inflammation, a higher-order regulation may control some liver-expressed genes that share a given chromosome area.
Subject(s)
Chromosomes/genetics , Gene Expression Regulation , Inflammation/genetics , Liver/metabolism , Liver/pathology , Physical Chromosome Mapping , Acute Disease , Animals , Cricetinae , Humans , Hybrid Cells , Liver/immunology , Mice , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Radiation Hybrid Mapping , Rats , Species SpecificityABSTRACT
PURPOSE: The work forms part of a major project to study the genetics of mouse cataract mutants found during the course of mutagenesis experiments. The long-term aim is to find the underlying gene mutation in each cataract mutant. Here we report further studies of the mutant cataract and curly whiskers (Ccw), previously mapped to Chromosome 4, and also investigations of the corneal opacity (Coop) mutant, which is shown to involve a mutation in the Pax6 gene. METHODS: For Ccw, the methods included mapping relative to microsatellite markers and histological studies. For the Coop mutant, breeding methods were used to show that Coop was allelic with Pax6. The Pax6 coding region in the mutant was then sequenced. RESULTS: The Ccw locus was mapped to approximately position 45cM on the consensus map of Chr 4. Histologically, progressive degeneration of the lens was seen. In the Coop mutant, a base-pair change C->T was found at position 1033 in the Pax6 gene, which created a stop codon leading to premature termination of translation, and to a truncated Pax6 protein. CONCLUSIONS: The phenotype in Ccw/+ heterozygotes involves a new type of lens degeneration in the mouse. On the basis of the phenotype and the locus position, no candidate gene has yet been identified. The Pax6coop mutant differs in phenotype from known null alleles of Pax6, implying that it is a hypomorph.
Subject(s)
Cataract/genetics , Corneal Opacity/genetics , DNA-Binding Proteins/genetics , Eye Diseases, Hereditary/genetics , Eye Proteins/genetics , Homeodomain Proteins , Vibrissae/abnormalities , Animals , Base Sequence , Cataract/pathology , Chromosome Mapping , Chromosomes/genetics , Consensus Sequence , Corneal Opacity/pathology , DNA Mutational Analysis , Eye Diseases, Hereditary/pathology , Female , Lens, Crystalline/pathology , Male , Mice , Mice, Inbred C3H , Mice, Inbred DBA , Mice, Mutant Strains , Microsatellite Repeats/genetics , Molecular Sequence Data , PAX6 Transcription Factor , Paired Box Transcription Factors , Repressor ProteinsSubject(s)
Chromosome Mapping , Evolution, Molecular , Genes/genetics , X Chromosome/genetics , Animals , Crosses, Genetic , DNA/genetics , Female , Haplotypes , Humans , Male , Mice , Microsatellite Repeats , Muridae , Polymorphism, Restriction Fragment LengthABSTRACT
The identification of many of the transcribed genes in man and mouse is being achieved by large scale sequencing of expressed sequence tags (ESTs). Attention is now being turned to elucidating gene function and many laboratories are looking to the mouse as a model system for this phase of the genome project. Mouse mutants have long been used as a means of investigating gene function and disease pathogenesis, and recently, several large mutagenesis programs have been initiated to fulfill the burgeoning demand of functional genomics research. Nevertheless, there is a substantial existing mouse mutant resource that can be used immediately. This review summarizes the available information about the loci encoding X-linked phenotypic mutants and variants, including 40 classical mutants and 40 that have arisen from gene targeting.
Subject(s)
Genetic Diseases, Inborn/genetics , Genetic Linkage , Mice/genetics , X Chromosome/genetics , Animals , Chromosome Mapping/methods , Humans , PhenotypeABSTRACT
We have demonstrated that the phenotype of the mouse mutant sex-linked fidget ( slf ) is caused by developmental malformations of the inner ear that result in hearing loss and vestibular dysfunction. Recently, pilot mapping experiments suggested that the mouse Brn4 / Pou3f4 gene co-segregated with the slf locus on the mouse X chromosome. These mapping data, in conjunction with the observation that the vertical head-shaking phenotype of slf mutants is identical to that observed in mice with a targeted deletion of the Brn4 gene, suggested that slf is a mutant allele of the Brn4 gene. In this paper, we have identified the nature of the slf mutation, and demonstrated that it is an X chromosomal inversion with one breakpoint close to Brn4. This inversion selectively eliminates the expression of the Brn4 gene in the developing inner ear, but not the neural tube. Finally, these results demonstrate that the slf mutation is a good mouse model for the most prevalent form of X-linked congenital deafness in man, which is associated with mutations in the human Brn4 ortholog, POU3F4.
Subject(s)
DNA-Binding Proteins , Ear, Inner/embryology , Ear, Inner/physiology , Mutation , Nerve Tissue Proteins , Transcription Factors/genetics , Animals , Chromosome Mapping , Female , Gene Expression Regulation, Developmental , Gene Rearrangement , Immunohistochemistry , Male , Mice , Mice, Inbred Strains , Mice, Knockout , Mice, Mutant Strains , POU Domain Factors , Reflex, Acoustic/physiology , Reflex, Startle/genetics , Regulatory Sequences, Nucleic Acid , Sex Factors , Transcription Factors/metabolism , X ChromosomeABSTRACT
Tattered (Td) is an X-linked, semi-dominant mouse mutation associated with prenatal male lethality. Heterozygous females are small and at 4-5 days of age develop patches of hyperkeratotic skin where no hair grows, resulting in a striping of the coat in adults. Craniofacial anomalies and twisted toes have also been observed in some affected females. A potential second allele of Td has also been described. The phenotype of Td is similar to that seen in heterozygous females with human X-linked dominant chondrodysplasia punctata (CDPX2, alternatively known as X-linked dominant Conradi-Hünermann-Happle syndrome) as well as another X-linked, semi-dominant mouse mutation, bare patches (Bpa). The Bpa gene has recently been identified and encodes a protein with homology to 3beta-hydroxysteroid dehydrogenases that functions in one of the later steps of cholesterol biosynthesis. CDPX2 patients display skin defects including linear or whorled atrophic and pigmentary lesions, striated hyperkeratosis, coarse lusterless hair and alopecia, cataracts and skeletal abnormalities including short stature, rhizomelic shortening of the limbs, epiphyseal stippling and craniofacial defects (MIM 302960). We have now identified the defect in Td mice as a single amino acid substitution in the delta8-delta7 sterol isomerase emopamil binding protein (Ebp; encoded by Ebp in mouse) and identified alterations in human EBP in seven unrelated CDPX2 patients.
Subject(s)
Chondrodysplasia Punctata/enzymology , Chondrodysplasia Punctata/genetics , Mutation , Steroid Isomerases/genetics , X Chromosome/genetics , Amino Acid Sequence , Animals , Base Sequence , Carrier Proteins/chemistry , Carrier Proteins/genetics , DNA Primers/genetics , Female , Genes, Dominant , Genetic Linkage , Guinea Pigs , Humans , Male , Mice , Mice, Mutant Strains , Molecular Sequence Data , Phenotype , Pregnancy , Sequence Homology, Amino Acid , Steroid Isomerases/chemistryABSTRACT
PURPOSE: The No2 cataractous mouse mutant displays a bilateral, congenital, hereditary nuclear opacity of the ocular lens. The aim of this work was to identify and subsequently screen an optimal candidate gene for a mutation correlated and consistent with the observed phenotype. METHODS: The No2 cataract was mapped in relation to genes and microsatellite markers by crossing to the wild mouse strain Mus spretus and then backcrossing to the inbred strain C3H/ HeH. The Cx50 (MP70) protein coding region and flanking sequences were amplified from normal parental as well as heterozygous and homozygous mutant genomic DNAs. These PCR products were then sequenced directly. Sequence data was corroborated by restriction analysis of PCR products. RESULTS: Mapping of the No2 cataract placed it in the vicinity of Gja8, the gene encoding connexin 50 (MP70), a major component of lens fiber gap junctions. Amplification and subsequent sequencing of the Cx50 protein coding regions revealed a single A-->C transversion within codon 47. This sequence change resulted in the creation of an HhaI restriction endonuclease restriction site, allowing for corroboration of the sequence data via restriction analysis using this enzyme. The sequence alteration is also predicted to result in the nonconservative substitution of alanine (Ala) for the normally encoded aspartic acid (Asp) at this position within the polypeptide. CONCLUSIONS: The identified mutation in Gja8 is both correlated and consistent with the cataract observed in the No2 mouse mutant, making it an ideal candidate for the cataract. This study provides the first evidence that a mutation in a lens connexin can result in congenital hereditary cataract, highlighting the importance of lens connexins in maintaining lens transparency.
Subject(s)
Cataract/genetics , Connexins/genetics , Eye Proteins/genetics , Lens Nucleus, Crystalline/pathology , Point Mutation , Animals , Cataract/pathology , DNA Primers/chemistry , Female , Male , Mice , Mice, Mutant Strains , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
MSG1 (melanocyte-specific gene 1) is a recently isolated gene predominantly expressed in cultured normal melanocytes and pigmented melanoma cells. MSG1 encodes a 27-kDa nuclear protein that has strong intrinsic transcriptional transactivating activity. In this report, the human MSG1 gene was mapped to chromosome Xq13.1 using X chromosome-specific somatic cell hybrids, and the mouse Msg1 gene was mapped 1.9 +/- 1.3 cM proximal to Xist using an interspecific backcross panel. Both the human and the mouse MSG1 genes consist of three exons and two introns within 5 kb of genomic DNA, and their genomic structures are highly conserved. Southern blot analysis suggests the existence of MSG1 homologues in chicken, zebrafish, and Drosophila. A 2.0-kb fragment of the 5'-flanking region of the mouse Msg1 gene contains a TATA box and potential binding sites for several transcription factors including USF, Brn-3, Brn-2, TFE3, Oct-1, AP-2, and Spl. This promoter fragment activates transcription of a reporter gene in pigmented melanoma cells, but not in amelanotic melanoma cells or nonmelanocytic cells, indicating that Msg1 expression is at least partially regulated at the transcriptional level.
Subject(s)
Chromosome Mapping , Nuclear Proteins/genetics , Promoter Regions, Genetic/genetics , X Chromosome/genetics , Animals , Apoptosis Regulatory Proteins , Base Sequence , Cloning, Molecular , Conserved Sequence , Exons/genetics , Genes, Reporter/genetics , Humans , Introns/genetics , Mice , Molecular Sequence Data , Sequence Analysis, DNA , TATA Box/genetics , Trans-Activators , Transcription Factors/genetics , Transcription, Genetic/genetics , Tumor Cells, CulturedABSTRACT
Patterning of the vertebrate limb along the anterior-posterior axis is controlled by the zone of polarizing activity (ZPA) located at the posterior limb margin. One of the vertebrate Hh family members, Shh, has been shown to be able to mediate the function of the ZPA. Several naturally occurring mouse mutations with the phenotype of preaxial polydactyly exhibit ectopic Shh expression at the anterior limb margin. In this study, we report the molecular characterization of a spontaneous mouse mutation, Doublefoot (Dbf). Dbf is a dominant mutation which maps to chromosome 1. Heterozygous and homozygous embryos display a severe polydactyly with 6 to 8 digits on each limb. We show here that Shh is expressed normally in Dbf mutants. In contrast, a second Hh family member, Indian hedgehog (Ihh) which maps close to Dbf, is ectopically expressed in the distal limb bud. Ectopic Ihh expression in the distal and anterior limb bud results in the ectopic activation of several genes associated with anterior-posterior and proximal-distal patterning (Fgf4, Hoxd13, Bmp2). In addition, specific components in the Hedgehog pathway are either ectopically activated (Ptc, Ptc-2, Gli1) or repressed (Gli2). We propose that misexpression of Ihh, and not a novel Smoothened ligand as recently suggested (Hayes et al., 1998), is responsible for the Dbf phenotype. We consider that Ihh has a similar activity to Shh when expressed in the early Shh-responsive limb bud. To determine whether Dbf maps to the Ihh locus, which is also on chromosome 1, we performed an interspecific backcross. These results demonstrate that Dbf and Ihh are genetically separated by approximately 1.3 centimorgans, suggesting that Dbf mutation may cause an exceptionally long-range disruption of Ihh regulation. Although this leads to ectopic activation of Ihh, normal expression of Ihh in the cartilaginous elements is retained.
Subject(s)
Polydactyly/genetics , Proteins/physiology , Trans-Activators , Transcription Factors , Transforming Growth Factor beta , Animals , Bone Morphogenetic Protein 2 , Bone Morphogenetic Proteins/genetics , Embryonic Induction/physiology , Extremities/growth & development , Extremities/pathology , Fibroblast Growth Factor 4 , Fibroblast Growth Factors/genetics , Gene Expression Regulation/genetics , Hedgehog Proteins , Homeodomain Proteins/genetics , Homozygote , In Situ Hybridization , Mice , Mice, Inbred Strains , Mutation/genetics , Proteins/genetics , Proto-Oncogene Proteins/genetics , Signal Transduction/physiologySubject(s)
Chromosome Mapping , Mice/genetics , X Chromosome/genetics , Animals , Chromosomes/genetics , Chromosomes, Human/genetics , Genome , Genome, Human , HumansABSTRACT
This article provides an introduction to genetic mapping for scientists who wish to map specific genes or mutant phenotypes in the mouse. The basic principles of genetic mapping and the different types of genetic markers available are described in the first two sections of the chapter. The theoretical and empirical principles necessary to consider when designing mapping experiments are reviewed in the third section. Protocols for mapping phenotypic traits and cloned genes are detailed in the fourth and fifth sections.
Subject(s)
Chromosome Mapping/methods , Genome , Animals , Chromosomes , Genetic Linkage/genetics , Genetic Markers/genetics , Genotype , Mice , Phenotype , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Recombination, Genetic/geneticsABSTRACT
The mouse X-linked mutants lined and stripey are associated with lethality of affected males in utero and a striping of the coat in carrier females. We demonstrate that the underlying mutations are nested deletions which lie in the Phex-Amelx chromosomal segment conserved between man and mouse. The lined deletion contains less than approximately 0.7 cM of genetic material and includes the growth factor-regulated protein kinase gene, Rsk2. Stripey carries a larger deletion which removes approximately 2.0 cM of genetic material, including Rsk2 and the pyruvate dehydrogenase E1alpha subunit gene, Pdha1 . Since Coffin-Lowry syndrome and neonatal lactic acidosis are associated with mutations in the human homologues of Rsk2 and Pdha1 respectively, lined and stripey provide models for gene deficiencies in these disorders.
Subject(s)
Abnormalities, Multiple/genetics , Acidosis, Lactic/genetics , Gene Deletion , Pyruvate Dehydrogenase Complex/genetics , Ribosomal Protein S6 Kinases/genetics , X Chromosome , Animals , Animals, Newborn , Body Weight , Chromosome Mapping , Crosses, Genetic , Female , Genetic Carrier Screening , Humans , Male , Mice , Mice, Inbred C3H , Mice, Mutant Strains , Muridae , Phenotype , Pyruvate Dehydrogenase Complex Deficiency Disease/genetics , Ribosomal Protein S6 Kinases/deficiencyABSTRACT
The genes for ocular albinisim type 1 (OA1) and the Xenopus laevis-like apical protein (APXL) map between amelogenin (AMELX) and the pseudoautosomal boundary in the distal region of the human X chromosome short arm. The mouse homologues, Oa1 and Apxl, have recently been shown to lie proximal to their expected locations on the mouse X chromosome, but their positions with respect to critical gene loci in the vicinity have not been defined. By analyzing recombination events from (Mus musculus x Mus spretus) x M. musculus backcrosses, we have constructed a detailed mouse genetic map that encompasses Oa1, five other genes, and 13 microsatellite loci. The order of genes and evolutionary breakpoints (EB) is defined as centromere-(EB)-(DXHXS674, DXHXS679)-Smcx-(EB)-Oa1-(EB)-Phex (3'-->5')-Pdha1-telomere. Thus Oa1 lies in a region between two previously characterized conserved segments.