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Silver fox gene mapping: conserved chromosome regions in the order Carnivora.

Twenty-three silver fox x hamster somatic cell hybrid clones were used to assign 15 fox genes: GPI to chromosome 1; PGD to chromosome 2; MDH2 to chromosome 3; ESD to chromosome 6; LDHB to chromosome 8; NP to chromosome 10; LDHA to chromosome 11; APRT, ENO1, and PGM1 to chromosome 12; IDH1 and MDH1 to chromosome 16; and GLA, G6PD, and HPRT to the X chromosome. High-resolution G-banding of human, cat, mink, and fox chromosomes containing homologous regions (according to genetic maps) revealed regions of putative homology. The results lend support to the suggestion that the most considerable karyotypic reorganization of the ancestral genome in the order Carnivora occurred during Canidae formation. The details of karyotypic evolution in mammals are discussed.

Animals↗

Why do stem cells exist?

Self-renewing tissues have a differentiation hierarchy such that the stem cells are the only permanent residents of the tissue, and it is in these cells that most cancerous mutations arise. The progeny of the stem cells either remain stem cells or enter a transient proliferating cell population that differentiates to produce the functional cells of the tissue. The reason that this differentiation hierarchy exists has not been established. We show here that alternative hierarchies, in which there would be no stem cells, are feasible and biologically plausible. We show that current evidence from somatic mutation frequencies at both transgenic and endogenous loci implicates cell division in the origin of most somatic mutations. We suggest, therefore, that the existence of stem cells is an evolutionary consequence of a selective pressure to avoid cancer by reducing the number of somatic mutations. The stem cell hierarchy reduces the number of cell divisions of those cells that reside permanently in the tissue, which reduces the number of somatic mutations and thus minimizes the cancer rate. In the small intestine, the existence of stem cells reduces the mutant frequency in the stem cells by about one order of magnitude. Since two or more mutations are required to transform a cell, the protective effect may be 100-fold or more. Similar factors may be expected in other tissues.

Age Factors↗

Identification and characterization of MTR1, a novel gene with homology to melastatin (MLSN1) and the trp gene family located in the BWS-WT2 critical region on chromosome 11p15.5 and showing allele-specific expression.

Alterations within human chromosomal region 11p15.5 are associated with the Beckwith-Wiedemann syndrome (BWS) and predisposition to a variety of neoplasias, including Wilms' tumors (WTs), rhabdoid tumors and rhabdomyosarcomas. To identify candidate genes for 11p15. 5-related diseases we compared human genomic sequence with expressed sequence tag and protein databases from different organisms to discover evolutionarily conserved sequences. Herein we describe the identification and characterization of a novel human transcript related to a putative Caenorhabditis elegans protein and the trp (transient receptor potential) gene. The highest homologies are observed with the human TRPC7 and with melastatin 1 ( MLSN1 ), whose transcript is downregulated in metastatic melanomas. Other genes related to and interacting with the trp family include the Grc gene, which codes for a growth factor-regulated channel protein, and PKD1/PKD2, involved in polycystic kidney disease. The novel gene presented here (named MTR1 for MLSN1 - and TRP -related gene 1) resides between TSSC4 and KvLQT1. MTR1 is expressed as a 4.5 kb transcript in a variety of fetal and adult tissues. The putative open reading frame is encoded in 24 exons, one of which is alternatively spliced leading to two possible proteins of 872 or 1165 amino acids with several predicted membrane-spanning domains in both versions. MTR1 transcripts are present in a large proportion of WTs and rhabdomyosarcomas. RT-PCR analysis of somatic cell hybrids harboring a single human chromosome 11 demonstrated exclusive expression of MTR1 in cell lines carrying a paternal chromosome 11, indicating allele-specific inactivation of the maternal copy by genomic imprinting.

Adult↗

Gene therapy treatments for erectile and bladder dysfunction.

The postgenomic age presents many exciting challenges and opportunities for the application of molecular medicine to the treatment of lower urinary tract diseases. Chief among these are the therapeutic possibilities afforded to selectively modulate/alter gene expression in somatic cells to "normalize" aberrant cellular responses to the existing hormonal milieu. In this paper, this therapeutic strategy will be referred to as gene therapy or gene transfer. This article specifically reviews the potential use of gene therapy/transfer to the treatment of bladder and erectile dysfunction. Available treatments for both of these common urologic diseases/disorders have contraindications, untoward side effects, or limited efficacy in certain patient populations. Moreover, that genetic material can be locally administered in the bladder and penis removes a major therapeutic obstacle to the use of gene transfer. Thus far, "proof-of-concept" has been demonstrated in preclinical studies using cellular and molecular strategies on a variety of gene targets. The continuing evolution of gene transfer vectors and gene delivery technologies is expected to further enhance the selectivity, efficacy, and duration of gene therapy, making it a viable treatment option for the amelioration of lower urinary tract diseases/disorders.

Erectile Dysfunction↗

Somatic excision of the Mu1 transposable element of maize.

The Mu transposons of the Robertsons's Mutator transposable element system in maize are unusual in many respects, when compared to the other known plant transposon systems. The excision of these elements occurs late in somatic tissues and very rarely in the germ line. Unlike the other plant transposons, there is no experimental evidence directly linking Mu element excision and integration. We have analyzed the excision products generated by a Mu1 transposon inserted into the bronze 1 locus of maize. We find that the excision products or 'footprints' left by the Mu1 element resemble those of the other plant transposable elements, rather than those of the animal transposable element systems. We also find some novel types of footprints resembling recombinational events. We suggest that the Mu1 element can promote intrachromosomal crossovers and conversions near its site of insertion, and that this may be another mechanism by which transposons can accelerate the evolution of genomes.

Base Sequence↗

Predominance of clonal reproduction, but recombinant origins of new genotypes in the free-floating aquatic bladderwort Utricularia australis f. tenuicaulis (Lentibulariaceae).

Aquatic plants are a biological group sharing several adaptations to aquatic conditions. The most striking evolutionary convergence in this group is the extensive reliance on clonal reproduction, which largely determines the patterns and process of evolution in aquatic plants. Utricularia australis f. tenuicaulis is a free-floating aquatic bladderwort that reproduces both sexually via seeds and clonally via turions and shoot fragments. Amplified fragment length polymorphism analysis was conducted on 267 ramets collected from 30 populations in Japan. The genotypic diversity within populations was extremely low, regardless of the geographical distribution range: the mean number of genotypes per population (G) was 1.4 and the mean genotypic diversity (D), including monoclonal populations, was 0.17. In contrast to the predominance of a few clones within populations, many of the populations investigated had different genotypes; a large portion of the genetic variation was explained by variation among populations. Character compatibility analysis clearly revealed that somatic mutations did not contribute to the origin of genotypic diversity in this aquatic bladderwort; instead, rare-to-sporadic sexual reproduction probably generated new genotypes. Thus, future studies should examine the role of sexual reproduction in this species from the viewpoint of long-term evolutionary benefits.

Ecosystem↗

Relationships among pathogenic and nonpathogenic isolates of Fusarium oxysporum based on the partial sequence of the intergenic spacer region of the ribosomal DNA.

Using PCR, we amplified and sequenced approximately 1,000 bp of the 5' end of the intergenic spacer (IGS) of the rDNA in 15 isolates of Fusarium oxysporum and one isolate of F. subglutinans. Isolates were selected to represent diversity in our collection based on differences in pathogenic race, vegetative compatibility group (VCG), mitochondrial DNA (mtDNA) haplotype, IGS haplotype, and DNA fingerprint. The objective of this research was to clarify the origin of virulence within F. oxysporum, the relationship between pathogenic and nonpathogenic strains, and the evolution of the different races of F. oxysporum f. sp. melonis. Bootstrapped parsimony analysis of the partial IGS sequence data identified a phylogenetic tree with highly significant branches. The two F. oxysporum f. sp. melonis VCGs, 0131 and 0134, were separated into distinct lineages. Race was not distinguished by significant IGS sequence differences within the pathogen VCGs. One exception was a race 1 isolate which was associated with VCG 0131 but, based on both mtDNA and IGS haplotype, had greater affinity with VCG 0134. Two IGS sequence types were found in this race 1 isolate, one suggesting an affiliation with VCG 0131 and the other similar to isolates in VCG 0134. This may have resulted from past somatic or sexual interactions between F. oxysporum f. sp. melonis, VCGs 0131 and 0134. Nonpathogens that were vegetatively compatible with the pathogen were not closely related to the pathogen based on IGS sequence data. Thus, nonpathogens and pathogens may share common alleles at vegetative compatibility loci by coincidence rather than because of recent clonal derivation from a common ancestor.

Base Sequence↗

Clonal evolution in Waldenstrom macroglobulinemia highlights functional role of B-cell receptor.

The course of clonal evolution of 2 related clones in the blood of a patient with Waldenstrom macroglobulinemia (WM) indicates the functional importance for the expression of the B-cell receptor for the survival of these malignant cells. Protein and nucleotide sequencing of the paraproteins' variable regions revealed 2 predominant Vlambda and 2 VH sequences, each set comprised in the ratio 1:1.5. The 2 VH sequences and 2 Vlambda sequences shared the same VDJ and VJ junctional sequences, respectively, indicating that 2 malignant clones had evolved from a common ancestor. This is the first report on intraclonal heterogeneity in WM. Comparison of the Vlambda and VH sequences with the closest matching known germline genes showed that they contained approximately 10 somatic mutations each. The distribution and type of mutations demonstrate that mutations have continued to accumulate in the malignant clones and that selection has been operating to preserve immunoglobulin structure.

Base Sequence↗

Structural characterization and mapping of the normal epithelial cell-specific 1 gene.

The normal epithelial cell-specific 1 (NES1) gene is a recently identified novel serine protease-like gene which is down-regulated during breast cancer progression. The gene product has 34-42% identity with the members of three distinct serine protease families: the trypsin-like family, activators of kringle domain-containing growth factors, and the kallikrein family (X. L. Liu et al., (1996) Cancer Res 56, 3371-3379). Although the cDNA of this gene has been cloned, its genomic structure and chromosomal position are not as yet known. Here, we report the genomic characterization and mapping of the NES1 gene. By subcloning and sequencing a PAC clone containing the complete NES1 gene, we were able to characterize the structure of this gene. The NES1 gene spans 5.5 kb and is composed of five coding exons and one untranslated exon. The positions of the introns were similar to trypsinogen, prostate specific antigen (PSA), and tissue plasminogen activator (TPA). NES1 gene was also localized with somatic cell mapping, radiation hybrid mapping, and fluorescence in situ hybridization techniques to chromosome 19q13.3-q13.4, the same region where the human kallikrein gene family resides. Taken together, our results suggest that the NES1 gene originates from the same ancestor as trypsinogen, PSA, and TPA, but remains in close proximity to PSA.

Amino Acid Sequence↗

Physical assignments of human chromosome 13 genes on pig chromosome 11 demonstrate extensive synteny and gene order conservation between pig and human.

Previous mapping between the human and pig genomes suggested extensive conservation of human chromosome 13 (HSA13) to pig chromosome 11 (SSC11). The objectives of this study were comparative gene mapping of pig homologs of HSA13 genes and examining gene order within this conserved synteny group by physical assignment of each locus. A detailed HSA13 to SSC11 comparison was chosen since the comparative gene map is not well developed for these chromosomes and a rearranged gene order within conserved synteny groups was observed from the comparison between HSA13 and bovine chromosome 12 (BTA12). Heterologous primers for PCR were designed and used to amplify pig homologous fragments. The pig fragments were sequenced to confirm the homology. Six pig STSs (FLT1, ESD, RB1, HTR2A, EDNRB, and F10) were physically mapped using a somatic cell hybrid panel to SSC11, and fluorescent in situ hybridization (FISH) mapping was also applied to improve map resolution and determine gene order. Results from this study increase the comparative information available on SSC11 and suggest a conserved gene order on SSC11 and HSA13, in contrast to human:bovine comparisons of this syntenic group.

Animals↗

Timing, genetic requirements and functional consequences of somatic hypermutation during B-cell development.

While somatic antibody mutants are rare in the preimmune repertoire and in primary immune responses, they dominate secondary and hyperimmune responses. We present evidence that somatic hypermutation is restricted to a particular pathway of B-cell differentiation in which distinct sets of B-cell clones are driven into the memory compartment. In accord with earlier results of McKean et al. (1984) and Rudikoff et al. (1984), somatic mutation occurs stepwise in the course of clonal expansion, before and after isotype switch, presumably at a rate close to 1 X 10(-3) per base pair per generation. At this rate, both selectable and unselectable mutations accumulate in the rearranged V region genes. The distribution of replacement mutations in the V regions shows that a fraction of the mutations in CDRs is positively selected whereas replacement mutations are counterselected in the FRs. By constructing an antibody mutant through site-specific mutagenesis we show that a point mutation in CDR1 of the heavy chain, found in most secondary anti-NP antibodies, is sufficient to increase NP binding affinity to the level typical for the secondary response. Somatic mutation may contribute to the immune repertoire in a more general sense than merely the diversification of a specific response. We have evidence that clones producing antibodies which no longer bind the immunizing antigen can be kept in the system and remain available for stimulation by a different antigen. Somatic mutations are 10 times less frequent in DJH loci than in either expressed or non-expressed rearranged VDJH or VJ loci. We therefore conclude that a V gene has to be brought into the proximity of the DJH segment in order to fully activate the hypermutational mechanism in these loci.

Animals↗

Comparative mapping using somatic cell hybrids.

Comparative mapping, or ascertaining the gene linkage relationships between different species, is rapidly developing. This is possible because new techniques in chromosome identification and somatic cell hybridization, such as the generation of hybrids preferentially segregating chromosomes of any desired species including rodents, and the development of gene transfer techniques have yielded new information about the human and rodent gene maps. In addition, the discovery and characterization of mouse subspecies has generated new mouse sexual genetic linkage data. The following picture is emerging. Several X-linked genes in man are X-linked in all mammalian species tested. The linkage relationships of several tightly linked genes, less than 1 map unit apart, are also conserved in all mammalian species tested. Ape autosomal genes are assigned to ape chromosomes homologous to their human counterparts indicating extensive conservation in the 12 million years (MYR) of evolution from apes to man. Similarly, mouse and rat, 10 MYR apart in evolution, have several large autosomal synteny groups conserved. In comparing the mouse and human gene maps we find that human genes assigned to different arms of the same human chromosome are unlinked in the mouse; mouse genes large map distances (20 to 45 cM) apart are very likely to be unlinked in the human. However, several autosomal synteny groups 10 to 20 cM apart, including the Pgd, Eno-1, Pgm-1 group on human chromosome arm 1p, are conserved in mice and man. This suggests that homology mapping, the superimposition of one species gene map on the homologous conserved portion of another species genome may be possible, and that ancestral autosomal synteny groups should be detectable.

Animals↗

[Do mitochondria play a role in aging?].

Ageing is an unavoidable and complex phenomenon which may be a price to pay to evolution. Thus genetics appear to play a predominant role besides environmental factors. Energetic metabolism slowly declines with ageing supporting a possible active role of mitochondria, the power supply of the cells, to this process. Mitochondrial DNA alterations appear during the mid-life and in degenerative diseases such as in Parkinson's and Alzheimer's; they include large scale deletions and point mutations. Since the respiratory chain plays a major role in the generation of superoxide anions which are converted into hydroxyl radicals that may impair lipids, proteins and DNA function in mitochondria, this vicious cycle may result from both an altered control of mitochondrial biogenesis dependent from the nucleus, and/or from a lack of repair and accumulation of somatic mitochondrial DNA mutations.

Aging↗

Nucleotide sequence of the promoter region of a tissue-specific human retroposon: comparison with its housekeeping progenitor.

The intronless autosomal phosphoglycerate kinase gene (Pgk-2) is a functional retroposon expressed in a tissue-specific manner in the meiotic and postmeiotic stages of mammalian spermatogenesis. The nucleotide sequence of the promoter region of this gene and its transcription start point are compared with those of Pgk-1, an intron-containing, X-linked, housekeeping gene expressed constitutively in all somatic cells and premeiotic germ cells. The location of flanking direct repeats and apparent conservation of specific regulatory sequences suggest the Pgk-2 retroposon arose from reverse transcriptase-mediated processing of an aberrant Pgk-1 transcript that included the endogenous Pgk-1 promoter elements. Specific sequences that may be involved in mediating differences observed in both the level and cell-type specificity of expression of these genes in spermatogenesis are identified.

Animals↗

Multigene family for sarcomeric myosin heavy chain in mouse and human DNA: localization on a single chromosome.

Cloned myosin heavy chain DNA probes from rat and human were hybridized to restriction endonuclease digests of genomic DNA from somatic cell hybrids and their parental cells. The mouse myosin heavy chain genes detectable by this assay were located on chromosome 11, and three different human sarcomeric myosin heavy chain genes were mapped to the short arm of chromosome 17. A synteny between myosin heavy chain and two unrelated markers, thymidine kinase and galactokinase, was found to be preserved in the rodent and human genomes.

Animals↗

Evolutionary dynamics of wAu-like Wolbachia variants in neotropical Drosophila spp.

Wolbachia bacteria are common intracellular symbionts of arthropods and have been extensively studied in Drosophila. Most research focuses on two Old Word hosts, Drosophila melanogaster and Drosophila simulans, and does not take into account that some of the Wolbachia associations in these species may have evolved only after their fast global expansion and after the exposure to Wolbachia of previously isolated habitats. Here we looked at Wolbachia of Neotropical Drosophila species. Seventy-one lines of 16 Neotropical Drosophila species sampled in different regions and at different time points were analyzed. Wolbachia is absent in lines of Drosophila willistoni collected before the 1970s, but more recent samples are infected with a strain designated wWil. Wolbachia is absent in all other species of the willistoni group. Polymorphic wWil-related strains were detected in some saltans group species, with D. septentriosaltans being coinfected with at least four variants. Based on wsp and ftsZ sequence data, wWil of D. willistoni is identical to wAu, a strain isolated from D. simulans, but can be discriminated when using a polymorphic minisatellite marker. In contrast to wAu, which infects both germ line and somatic tissues of D. simulans, wWil is found exclusively in the primordial germ line cells of D. willistoni embryos. We report on a pool of closely related Wolbachia strains in Neotropical Drosophila species as a potential source for the wAu strain in D. simulans. Possible evolutionary scenarios reconstructing the infection history of wAu-like Wolbachia in Neotropical Drosophila species and the Old World species D. simulans are discussed.

Animals↗

A family of IGF-II mRNA binding proteins (IMP) involved in RNA trafficking.

During a search for trans-acting factors associating with insulin-like growth factor II (IGF-II) mRNAs, we recently identified a family of three IGF-II mRNA-binding Proteins (IMP1, IMP2 and IMP3) that exhibit multiple attachments to IGF-II leader 3 mRNA and the reciprocally imprinted H19 RNA. IMPs contain the unique combination of two RNA recognition motifs (RRMs) and four hnRNP K homology (KH) domains. IMP1 is orthologous to the chicken zipcode-binding protein (ZBP-1), and the mouse c-myc coding region determinant-binding protein (CRD-BP) that associates with beta-actin and c-myc mRNA, respectively. Moreover, the p62 protein identified in hepatocellular carcinoma represents a splice variant of IMP2, and IMP3 is orthologous to the Xenopus Vegetal 1 RNA-binding protein (Vgl-RBP/Vera). IMPs are produced in a biphasic fashion--initially during the early stages of embryogenesis and subsequently later in development. IMPs and their orthologues are predominantly cytoplasmic and are implicated in the transport of their RNA targets towards the leading edge in somatic cells and to the vegetal pole in Xenopus oocytes, respectively. RNA localization is a conserved mechanism of polarizing genetic information in the establishment of asymmetries during both embryogenesis and adult life, enabling local protein synthesis at final destinations within the cell. The identification of developmentally expressed zipcode-binding proteins indicates that RNA trafficking participates in processes such as cell-growth and migration during embryogenesis.

Amino Acid Sequence↗

[Psychiatric consultation in pediatrics. Description and problems].

In this study of psychiatric consultations in a paediatric department, a description is given of the evolution and duration of treatment in the first 55 cases seen. Children ranged from age 1 month to 12 years. Two findings are of particular interest: most of the children were of a very young age (less than 3 years); most psychiatric treatments were of short duration. These results are due to the preventive orientation of our consultation and to the close collaboration between pediatricians and psychiatrists. The preventive approach allows for less heavy and costly measures than the standard long-term psychiatric treatment. This preventive approach is only possible if the diagnosis is established early. The pediatrician is in a better position than the psychiatrist to practice early detection, as in young children the psychopathology is often expressed through somatic channels. In cases where the pathology was detected at a later age, the treatment is usually more difficult.

Age Factors↗