Chromosome assignment of the gene loci ETS1 and THY1 in the owl monkey

Our previous assignment of the gene loci HBB, HRAS1, INS, PTH, LDHA, and CAT to owl monkey chromosome 19 of karyotype VI (K-VI) indicated a putative homology of this owl monkey chromosome with the short arm of human chromosome 11 (HSA 11p). To investigate further the extent of shared homology, we localized in the owl monkey complement two genes known to be on HSA 11q. Segregation analysis of ETS1 and THY1 homologous DNA in three karyotypically different panels of rodent x owl monkey somatic cell hybrids provided evidence for the syntenic assignment of these loci to homologous chromosomes of three owl monkey karyotypes, namely, chromosomes 4 (K-VI), 3 (K-II), and 5 (K-V). The results indicate a disruption of syntenic gene loci on the distal portion of HSA 11q from 11p during primate evolution.     

A putative homoeolog of human chromosome 12 in the owl monkey

Analyses of Southern blots of rodent x owl monkey somatic cell hybrids permitted syntenic assignment of gene loci coding for triosephosphate isomerase (TPI), antigen CD4(T4), Kirsten rat sarcoma 2(KRAS2) virus, insulin-like growth factor 1 (IGF1), and alpha 2-macroglobulin (A2M) to chromosome 10 of owl monkey karyotype VI(2n = 49, 50). In addition, regional in situ localization of the T4 and KRAS2 loci on the proximal region of the long arm of this acrocentric chromosome and on the corresponding homologous region on the long arm of metacentric chromosome 1 of karyotype IV (2n = 52) substantiated our hypothesis that a single fusion or fission event is responsible for the polymorphism in chromosome number characteristic of owl monkeys from at least three allopatric populations. The study supports a putative homoeology between owl monkey chromosome 10 (K-VI) and human chromosome 12. The morphological differences between these two primate chromosomes indicate evolutionary rearrangements involving at least one pericentric inversion.     

Mapping of five human chromosome 19 DNA markers to owl monkey chromosomes.

Hybridization of DNA from three panels of karyotypically distinct owl monkey x rodent somatic cell hybrids with human DNA probes resulted in the syntenic assignments of INSR-LDLR-TGFB1-APOE-D19S8 to owl monkey chromosome 25 of karyotype VI (2n = 49/50), INSR-LDLR-TGFB1-D19S8 to chromosome 2 of karyotype II (2n = 54), and INSR-APOE to chromosome 2 of karyotype V (2n = 46). The APOE and D19S8 loci are on adjacent regions proximal to the centromere of chromosomes 25q (K-VI) and 2p (K-II), as determined by in situ chromosomal hybridization analysis. These findings support our previous proposals on (1) the homology of these chromosomes of three owl monkey karyotypes, (2) the evolutionary derivation of chromosome 2 of karyotypes II and V as the result of two separate centric fusion events, and (3) the likelihood that owl monkey chromosome 25 (K-VI) (and its homologs) is a conserved genetic homoeolog of human chromosome 19.     

Comparative chromosomal mapping of the owl monkey serum albumin gene

We have mapped the albumin locus (ALB) in the owl monkey, Aotus trivirgatus, using a cloned human albumin gene probe, pcHSA 33-1. Rodent-owl monkey somatic cell hybrids were used to map the owl monkey albumin locus in three subgroups of Aotus, karyotypes II, V, and VI. Segregation analysis of the molecular hybridization pattern of pcHSA 33-1 in the somatic cell hybrids indicated that the albumin locus maps to chromosome 9 of owl monkey karyotype II, chromosome 12 of karyotype V, and chromosome 1 of karyotype VI. This assignment provides evidence for the homology of these three chromosomes and supports the hypothesis of Ma on the formation of chromosome 1 in karyotype VI.     

Molecular evidence of Y-autosomal translocations in owl monkeys

Probe pDP1007, which contains highly conserved DNA sequences from the sex-determining region of the human Y chromosome, cross-hybridized with owl monkey EcoRI restriction fragments of 1.8 kb and 6.6 kb. Southern transfer analysis of owl monkey (karyotype VI)--rodent somatic cell hybrids localized the 1.8-kb fragment on the owl monkey X chromosome and the 6.6-kb fragment, which is male specific, on chromosome 14/Y. Regional in situ chromosome mapping of pDP1007 revealed specific sites of hybridization: the distal short arm of the X chromosome of karyotypes IV, VI, and VII; the small metacentric Y of karyotype IV; the C-band positive region on the short arm of chromosome 17/Y (karyotype VII); and the C-band positive region on the long arm of chromosome 14/Y (karyotype VI). These molecular findings reinforce cytological evidence that Y-chromosomal material has been transferred to autosomes 14 and 17 in owl monkeys of karyotypes VI and VII, respectively, in which there are no independently segregating Y chromosomes.     

Homologs of eleven loci on human chromosome 11 assigned to two owl monkey chromosomes

We localized 11 loci mapped to human chromosome 11 to two chromosomes, 4 and 19 of owl monkey karyotype VI (2n = 49/50), by the use of somatic cell hybrids. Furthermore, using in situ hybridization to chromosomes of two owl monkey karyotypes, the HSTF1 oncogene locus was precisely localized on homologs 19q (K-VI) and 2q (K-II). Comparative analysis of available gene-mapping data among human, mouse, and owl monkey chromosomes revealed a pattern of evolutionary change in a syntenic group on human chromosome 11. These structural changes could be explained as having derived from a pericentric inversion of human chromosome region 11cen----q13 and a translocation involving human region 11q22----qter during primate evolution.     

Owl monkey gene loci for c-fes and albumin are syntenic

Segregation analysis of rodent-owl monkey somatic cell hybrids probed with a human c-fes DNA sequence disclosed the owl monkey c-fes locus to be syntenic with the albumin loci on chromosome 1 of karyotype VI, chromosome 12 of karyotype V, and chromosome 9 of karyotype IV. This finding substantiates the proposed homology of these chromosomes and demonstrates the complexity of rearrangement of individual genes during chromosomal phylogeny which is not detectable by morphological comparison alone.     

Chromosome localization and gene synteny of the major histocompatibility complex in the owl monkey,Aotus

Rodent cells were hybridized with owl monkey (Aotus) cells of karyotypes II, III, V, and VI. Aotus-rodent somatic hybrid lines preferentially segregating Aotus chromosomes were selected to determine the chromosomal location of the major histocompatibility complex and other genes with which it is syntenic in man. Based on correlation between concordant segregation of the chromosome as visualized by G-banding and expression of the Aotus antigens or enzymes in independent Aotus-rodent hybrid clones, we have assigned Aotus gene loci for the MHC, GLO, ME₁, SOD₂, and PGM₃ to Aotus chromosome 9 of karyotype VI (2n=49/50), chromosome 10 of karyotype V (2n=46), and chromosome 7 of karyotypes II and III (2n = 54 and 53). On the basis of banding patterns we previously postulated that these chromosomes of the different karyotypes were homologous. The gene assignments reported here provide independent evidence for that hypothesis. Aotus chromosomes 9 (K-VI), 10 (K-V), and 7 (K-II, III) are homologous to human chromosome 6 in that they all code for the MHC, GLO, ME₁, SOD₂, and PGM₃. The structural differences between these homologous chromosomes probably resulted from a pericentric inversion.Abbreviations used in this paper MHC major histocompatibility complex - HLA human lymphocyte antigen - PGM3 phosphoglucomutase-3 - ME₁ cytoplasmic malic enzyme-1 - SOD₂ superoxide dismutase-B - GLO glyoxalase 1 - OMLA owl monkey leukocyte antigens - K karyotype - ₂-M ₂-microglobulin - DMEM Dulbecco's modification of Eagle's medium - PEG polyethylene glycol - HAT hypoxanthine, aminopterin, and thymidine - KC1 potassium chloride - G-band-trypsin Giemsa band     

In vitro selection of SV40 T antigen epitope loss variants by site-specific cytotoxic T lymphocyte clones

SV40-transformed cells of C57BL/6 (B6) mouse origin (H-2b) express four distinct predominant antigenic sites, I, II, III, and IV, on SV40 large tumor (T) Ag that are recognized by SV40 T Ag-specific CTL clones. In this study, we selected SV40 T Ag-positive cell lines which had lost one or more of the antigenic sites, by in vitro cocultivation of a SV40-transformed B6 mouse kidney cell line (K-0) with SV40 T Ag site-specific CTL clones, Y-1 (site I specific), Y-2 (site II specific), Y-3 (site III specific), and Y-4 (site IV specific). All of the CTL-resistant cell lines expressed large quantities of cell surface H-2 class I Ag. K-1 cells selected by CTL clone Y-1 lost the expression of antigenic sites I, II, and III, but not site IV. K-2 and K-3 cells selected by CTL clones Y-2 and Y-3, respectively, were found to be negative for sites II and III but expressed sites I and IV. K-4 cells selected by CTL clone Y-4 lost the expression of only site IV. K-1,4 cells (sites I-, II-, III-, IV-) were selected from K-1 cells by cocultivation with CTL clone Y-4, K-2,4 cells (sites I+, II-, III-, IV-) were selected from K-2 cells by CTL clone Y-4. K-3,1 cells (sites I-, II-, III-, IV+) were selected from K-3 cells by CTL clone Y-1, and K-3,1,4 cells (sites I-, II-, III-, IV-) were selected from K-3,1 cells by CTL clone Y-4. From K-4 cells, K-4,1 cells (sites I-, II-, III-, IV-) and K-4,3 cells (sites I+, II-, III-, IV-) were selected by CTL clone Y-1 and Y-3, respectively. The antigenic site loss variant cell lines K-1, K-1,4, K-3,1 K-3,1,4, K-4,1, and K-4,3 synthesized SV40 T Ag molecules of 75, 75, 78, 78, 81, and 88 kDa, respectively. Expression of wild-type SV40 T Ag in the antigenic site loss variants by infection with SV40 or transfection with cloned SV40 DNA restored the CTL recognition sites on the variant cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Two new karyotypes in the Peruvian owl monkey (Aotus trivirgatus)

The observation of remarkable karyotypic variation in owl monkeys (Aotus trivirgatus) stimulated us to study the chromosomal evolution of this New World genus. As an extension of this project, we examined the chromosome complement of a “phenotype-B” Aotus population from Peru. In addition to karyotype V(2n = 46), two new karyotypes with diploid numbers of 47 and 48 were identified. A G-band comparison of these karyotypes indicated that the chromosome number polymorphism in these Peruvian owl monkeys resulted from a single fusion or fission event involving a single metacentric and two acrocentric chromosome pairs. This mechanism is also known to be responsible for the chromosome number polymorphism in at least two other populations of phenotype B Aotus, one from Colombia and the other from Panama.     

Owl monkey gene mapping: the assignment of gene loci for catalase, beta-globin gene cluster, HRAS1, insulin, and parathyroid hormone

Using somatic cell genetics and Southern blot hybridization, we have mapped five structural genes in the owl monkey, coding for catalase (CAT), the beta-globin gene cluster (HBBC), c-Ha-ras 1 (HRAS1), insulin (INS), and parathyroid hormone (PTH). All five loci are mapped to chromosome 19 of karyotype VI (2n = 49,50) of the owl monkey; CAT, HBBC, INS, and PTH can be assigned to chromosome 4 of karyotype V (2n = 46), while CAT and HBBC can be assigned to chromosome 2 of karyotype III (2n = 53). Using in situ hybridization, the CAT gene was precisely mapped on the mid-region and the beta-globin gene cluster on the telomeric end of chromosome 2q(K-III). Our results provide significant insight into the evolutionary history of these gene loci. While these loci are separated into at least two major segments in rodents such as the mouse, our results suggest conservation of a single chromosome arm among higher primates.     

Anchoring of genetic linkage maps to the chromosome complement of Vicia faba L.

Using amplification of marker sequences with DNA from a set of distinct microdissected Vicia faba L. chromosomes covering the entire genome, we could unambiguously show that the linkage group I.B, which includes the pseudogene of legumin B4 (ψ1) and was previously ascribed to the metacentric chromosome I, actually belongs to chromosome IV. By considering the breakpoints of the translocated BKH chromosomes III and IV, even the subchromosomal position of loci LG085 and CNGC4 could be inferred. Anchoring all linkage groups to distinct faba bean chromosomes will facilitate quantitative trait locus fine mapping and gene identification using synteny, and will boost the development of efficient markers for selection in breeding programs.     

Karyotypic analysis of two algae species Scenedesmus incrassatulus Bohl and Scenedesmus antennatus Bréb (Chlorophyta, Chlorococcales)

The karyotypes (number, morphology and size of the chromosomes) of two algae species of Scenedesmus genus, S. incrassatulus and S. antennatus, were studied. The karyotype of S. incrassatulus (n=4) was asymmetric, characterized by two large metacentric, one large submetacentric and one small metacentric chromosomes. The karyotype assembly of S. antennatus (n=6) reveals two metacentrics and four submetacentrics. This karyotype was symmetric. The general chromosomal formulae of both species, as well as the total average metaphase length of their haploid set are presented. The results of chromosomal studies of other related species are compared and discussed. Data from the karyotypic analysis showed that S. incrassatulus, S. antennatus and S. obliquus are separate biological species from taxonomical point of view.     

Chromosome assignment of the owl monkey MOS and MYC gene loci

Southern blot hybridizations of rodent x owl monkey hybrid DNAs with human cDNA probes allowed the mapping of the MOS and MYC gene loci to owl monkey chromosome 16 of karyotype VI (2n = 49 male/50 female) and to the homologous chromosome 15 of karyotype V (2n = 46). Synteny of MOS and MYC gene loci in both man and owl monkey suggests this chromosome segment's conservation in primates, contrasting with its disruption in the mouse.     

Genomic organization of the yeast Yarrowia lipolytica

We produced electrophoretic karyotypes of the reference strain E150 and of seven other isolates from different geographical origins to study the genomic organization of the dimorphic yeast Yarrowia lipolytica. These karyotypes differed in the number and size of the chromosomal bands. The karyotype of the reference stain E150 consisted of five bands of between 2.6 and 4.9 Mb in size. This strain contained at least five rDNA clusters, from 190 to 620 kb in size, which were scattered over most of the chromosomes. The assignment of 43 markers, including rRNA genes and three centromeres, to the E150 bands defined five linkage groups. Hybridization to the karyotypes of other isolates with pools of markers of each linkage group showed that linkage groups I, II, IV and V were conserved in the strains tested whereas group III was not and was split between at least two chromosomes in most strains. Use of a meganuclease I-SceI site targeted to one locus of E150 linkage group III showed that two chromosomes actually comigrated in band III of this strain. Our results are compatible with six chromosomes defining the haploid complement of strains of Y. lipolytica and that, despite an unprecedented chromosome length polymorphism, the overall structure of the genome is conserved in different isolates. Received: 27 March 1997; in revised form: 8 July 1997 / Accepted: 9 July 1997     

Karyotype of the Snow Sculpin <Emphasis Type="Italic">Myoxocephalus brandti</Emphasis> Steindachner (Cottidae) from Peter the Great Bay,Sea of Japan

The karyotype of the snow sculpin Myoxocephalus brandti, 2n = 44, NF = 46, from Peter the Great Bay was studied. Two-armed chromosomes were presented by one pair of metacentric chromosomes of medium size; one-armed chromosomes included two pairs of large subtelocentric chromosomes and a pair of large acrocentric chromosomes. Ag-NOR-staining in the telomere vicinity revealed nucleolus-organizing regions in one metacentric chromosome and in one medium size acrocentric chromosome in one of the fishes, in two homological small acrocentric chromosomes in three fishes, and in one acrocentric chromosome of average size in six fishes. No difference between the male and female karyotypes and any type of variability was revealed. The karyotypes of the snow sculpin M. brandti and the frog sculpin M. stelleri were compared. Their distinctions and similarities were displayed.Original Russian Text Copyright ¢ 2005 by Biologiya Morya, Ryazanova.     

Chromosomes of the liver fluke, Clonorchis sinensis

A karyological study was carried out in order to compared the chromosome numbers, chromosome morphologies and karyotypes of the oriental liver fluke, Clonorchis sinensis (Trematoda: Opisthorchiidae), collected from Korea and China. Chromosome preparations were made by means of air-drying method. The chromosome number was 2n = 56 in both Korean and Chinese flukes, and chromosomes were divided into two groups based on this size; consisting of 8 pairs of large and 20 pairs of small chromosomes. However, the karyotypes showed some differences between Korean and Chinese flukes. The karyotype of liver flukes from Korea consisted of three metacentric pairs, one meta-/submetacentric pair, 16 submetacentric pairs and eight subtelocentric pairs of chromosomes. On the other hand, liver flukes from China consisted of two metacentric pairs, two meta-/submetacentric pairs, 16 submetacentric pairs and eight subtelocentric pairs of chromosomes.     

Linkage-dependent gene flow in a house mouse chromosomal hybrid zone

In the alpine valley of Valtellina there are two Robertsonian chromosomal races of house mouse, the Poschiavo (POS: 2n = 24-26) characterized by metacentric 8.12 and acrocentrics 2 and 10 and the Upper Valtellina (UV: 2n = 22-24) characterized by metacentrics 2.8 and 10.12. The races inhabit separate villages in the valley except in Sommacologna and Sondalo, where they both occur together with hybrids. A total of 179 mice from 16 villages were typed at 13 microsatellite loci. Seven of these loci were localized close to the centromeres of chromosomes 10 and 12, with the prediction that these regions on the race-specific chromosomes would be the most likely to experience a barrier to gene flow. The remaining six loci were localized at the telomeres of chromosomes 10 and 12 and at the centromeres of chromosomes that do not differ between the races. Substantial differences in allelic frequencies were found between the villages with POS and UV races at five of the loci at the centromeres of chromosomes 10 and 12 but at none of the other loci. These differences were not found to distinguish the two races in Sommacologna and Sondalo. Therefore, the centromeric regions of race-specific chromosomes do appear to experience a barrier to gene flow, although this can break down under intense interbreeding between the races. These results are considered in the context of Harrison's (1990) concept of the semipermeability of hybrid zones to gene exchange and in relation to parapatric speciation.     

A primate transfer RNA gene cluster and the evolution of human chromosome 1.

The localisation of tRNA(Asn) gene clusters in the karyotypes of primates has been studied by means of in situ hybridisation. In the human and orangutan (Pongo pygmaeus) karyotypes there are two such gene clusters, one each on the long and short arms of chromosome 1. Old World monkeys, however, contain both gene clusters on their equivalent of the human chromosome 1 short arm, which can be explained by a pericentric inversion which (amongst other chromosome changes) distinguishes the human and Old World monkey chromosomes 1. The capuchin (Cebus appella), however, a New World monkey, has only one tRNA(Asn) gene cluster, at least on the elements equivalent to human chromosome 1. This cluster is located proximal to the centromere on a chromosome that has been tentatively identified (by others) as the equivalent of the long arm of human chromosome 1. Should this prove to be correct, it would indicate that the large primate metacentric came into being in the form found today in the great apes, rather than in the form currently found in Old World monkeys. These data further show that the tRNA(Asn) gene cluster has been split in two since before the Old World monkeys and hominids diverged, i.e., over 30 million years ago, and also that the original transfer of these genes from one arm of chromosome 1 to the other was unlikely to have involved a pericentric inversion but, rather, some form of replicative transposition.