Replication studies on prometaphase chromosomes of man, the chimpanzee, the pygmy chimpanzee, the gorilla, and the orangutan reveal great interspeeifie homologies between the autosomes. The early replicating X chromosomes clearly show a high degree of conservation of both the pattern and the time course of replication. An early replicating segment on the short arm of the X chromosomes of man (Xp22.3) which escapes inactivation can be found on the X chromosomes of the great apes as well. Furthermore, the most early replicating segment on the Y chromosomes of all species tested appears tö be homologous to this segment on the X chromosomes. Therefore, these early replicating segments in the great apes may correspond to the pseudoautosomal segment proposed to exist in man. From further cytogenetic characterization of the Y chromosomes it is evident that structural alterations have resulted in an extreme divergence in both the euchromatic and heterochromatic parts. It is assumed, therefore, that, in contrast to the X chromosomes, the Y chromosomes have undergone a rapid evolution within the higher primates. Despite numerous comparative cytogenetic studies in man and the great apes (see, e.g., Turleau et al., 1972; Dutrillaux, 1975; Egozcue, 1975; Schnedl et al., 1975; Miller, 1977; Seuänez, 1979; Schmidetal . , 1981; Yunis and Prakash, 1982), the Y and the late-replicating X chromosomes are only poorly characterized in the great apes. Replication studies on prometaphase human sex chromosomes have revealed a distinet early replicating segment on the distal ends of both Xp and Yp (Müller and Schempp, 1982; Schempp and Müller, 1982), corresponding to the pairing segment in which a synaptonemal complex is formed during male meiosis (Moses et al., 1975; Solari, 1980; Chandley et al., 1984). Furthermore, this early replicating segment on Xp appears to escape inactivation (Race and Sanger, 1975; Mohandas et al., 1979; Müller et al., 1980; Wolf, 1981); therefore, it was suggested that the segSupported by the Deutsche Forschungsgemeinschaft (Schempp 214/3-1). Request reprints f r o m : Dr. W. Schempp, Institut für Humangenetik und Anthropologie, Universität Freiburg, Albertstrassc 11, D-7800 Freiburg i. Br. (Fedcral Republic of Germany). ments on the distal portions of Xp and Yp are a remnant left unchanged during the differentiation of heteromorphic sex chromosomes from originally homomorphic autosomes (Schempp and Meer, 1983). The question arises whether the sex chromosomes of the great apes demonstrate a similar replication behavior. Therefore, we have performed a comparative replication study on prometaphase chromosomes, paying special attention to the sex chromosomes of man, the chimpanzee, the pygmy chimpanzee, the gorilla, and the orangutan. In order to draw a comprehensive comparison of the Y chromosomes of man and the great apes, additional staining procedures for the characterization of heterochromatin were applied. Materials and methods C h r o m o s o m e p r e p a r a t i o n s Chromosome preparations from five normal human males and fcmalcs ( H o m o sapiens [HSA]), five male and three female chimpanzees ( P a n t r o g l o d y t e s [PTR], T N O Primate Center, Rijswijk, The Nethcrlands; Tierpark Hellabrunn, München, FRG; Zoologischer Scx-chromosomc replication in man and the great apes 73 Garten, Frankfurt, FRG; Zoologischer Garten, Berlin, FRG) , two male pygmy chimpanzees ( P a n paniscus [PPA], Zoologischer Garten, Köln, FRG), three male and two female gorillas ( G o r i l l a g o r i l l a [GGO], Zoologischer Garten, Köln, FRG; Zoologischer Garten, Berlin, FRG), and four male and three female Bornean orangutans ( P o n g o pygmaeus [PPY], Zoologischer Garten, Köln, FRG; Tierpark Hellabrunn, München, FRG) were made from peripheral lymphocytes aecording to the method of Pfeiffer (1974), with minor modifications (Schempp and Meer, 1983). Briefly, the cells were cultivated in RPMI 1640 (Gibco), supplemcnted with 15% fetal calf serum and phytohemagglutinin. After 65 h of ineubation, bromodeoxyuridine (BrdU, 10 ug/ml) and fluorodeoxyuridine (FdU, 0.5 ug/ml) were added; 6 h later thecultures were treated with Colcemid (0.05 ug/ml) for 30 min. The cells were then harvested by centrifugation, resuspended in hypotonic Solution (0.4% KCl) for 30 min at 37 C, and fixed in 3:1 methanol:acetic acid. Cells were washed three times with fixative and kept at 4°C overnight. Chromosomes were spread on cold slides by flaming. Slides were air-dried for al least 24 h. S t a i n i n g methods B r d V r e p l i c a t i o n p a t t e r n s . Prometaphase chromosome preparations of the BrdU-lreated cultures were differentially stained with acridine orange (50 ug/ml), resulting in RBA patterns (ISCN, 1985). By adding BrdU to the cultures during the last 6.5 h, thymidine is incorporated inlo early replicating chromosome sites, resulting in bright fluorescence (light bands), while the incorporation of BrdU into late-replicating chromosome material produces dull-red staining (dark bands). Q b a n d i n g . Quinacrine mustard was used aecording to the technique of Caspersson et al. (1970). D i s t a m y c i n A / D A P I b a n d i n g . A slight modification of the technique originally described by Schweizer et al. (1978) was applied. In brief, the slides were ineubated in distamycin A (50 ug/ml in Mcllvainc's buffer, pH 7.0) for 20 min at room lemperature. The slides were then lightly washed in buffer and stained with DAPI (1 ug/ml in Mcllvaine's buffer, pH 7.0) for a furlher 20 min. After a rinse in buffer, the preparations were mounted in equal parls of glycerol and Mcllvaine's. For fluorescence microscopy, a BP 365/FT 395/LP397 (Zeiss) filter combination was used. C b a n d i n g . Constitutive heterochromatin was stained aecording to the method of Sumner (1972). C h r o m o s o m e length measurements Centromere indices of the Y chromosomes of man and the great apes were determined by length measurements of 30 Q-banded metaphase plates of each species. For interspeeifie comparison of the Y chromosomes, the X-chromosome length was used as an
By fluorescence in situ hybridization (FISH), we constructed a chromosome map of 324 site-specific clones and 119 painting clones, which were isolated from a cosmid library of Chinese hamster genomic DNA. The site-specific clones were distributed among all chromosomes except chromosome 10 and the Y chromosome. On the other hand, FISH signals of painting clones were mainly found in heterochromatic regions. First, the various combinations of clones painting centromeric regions suggest that the centromeric heterochromatin of Chinese hamster chromosomes is extremely diverse in its repetitive sequence composition. Second, the long arm of the X chromosome and the whole Y chromosome, both heterochromatic regions, were simultaneously painted by many clones. The pattern of co-localization indicated that a proximal segment of the long arm of the X chromosome matches the complete long arm of the Y chromosome, whereas a distal segment of the long arm of the X chromosome is comparable to the whole short arm of the Y chromosome. Thus, these findings suggest that the corresponding segments of the X and Y-chromosomes have common repeated DNA elements.
Regulated inhibition of programmed cell death (apoptosis) may control tissue differentiation and aberrantly promote cell survival in cancer. The extensive work in the field of apoptosis during the last decade has resulted in the identification of several “inhibitor of apoptosis proteins” (IAPs), which are highly conserved from virus to mammals. Survivin (BIRC5, baculoviral IAP repeat-containing 5) is the smallest member of the IAP gene family and it has been isolated in human (Ambrosini et al., 1997) and mouse (Li and Altieri, 1999). It maps to 17q25 and 11E2 in the two species, respectively. The complementary strand of the BIRC5 gene codes for the effector cell protease receptor-1 (EPR1) (Altieri, 1995). Southern blots of genomic DNA have pointed to the presence of multiple BIRC5/EPR1related genes, which are evolutionarily conserved. Northern blot studies have demonstrated the existence of two independent transcripts for BIRC5 and EPR1 of 1.9-kb and 1.3-kb, respectively (Ambrosini et al., 1998). BIRC5 is expressed in the G2/M phase of the cell cycle and is associated with mitotic spindle microtubules. Disruption of these interactions causes the loss of the anti-apoptosis function, resulting in an aberrant progression of transformed cells (neoplasia) (Li et al., 1998). It has been suggested that a better understanding of the molecular mechanisms of the action of BIRC5 may help in the manipulation of the sensitivity of cancer cells to therapy. The BIRC5 transcript is found in fetal tissues and in transformed cell lines in the most common human cancers in vivo (Ambrosini et al., 1997). EPR1 is involved in signal transduction in the coagulation process and is expressed in normal adult and fetal tissues (Ambrosini et al., 1998). In the present study we report the isolation of one clone from a pig small intestine cDNA library containing the BIRC5 and the EPR1 genes. The entire clone has been sequenced and the sequence has been used to search for homologies in the databases. This clone has been mapped in pig using somatic and radiation hybrid panels.
A second human orthologue of a new, putative rat brain Lglutamate (Glu) receptor-like gene, GRINL1B, has been isolated. An intron-containing version of this gene, GRINL1A, has been previously isolated and mapped to human chromosome 15q22.1 (Roginski et al., 2001). The GRINL1B genomic sequence is nearly identical to the GRINL1A cDNA sequence (AF326773); that is, the GRINL1B gene is devoid of introns. However, GRINL1B may encode gene products (see Results and discussion). Recent studies (Cook et al., 1998; IMGSAC, 1998; Philippe et al., 1999; Risch et al., 1999) reported probability peaks for autism related genes in the region where we mapped the GRINL1A gene. We therefore mapped the GRINL1B gene to determine whether it also might be located in an autism susceptibility region. Materials and methods
Alterations in nuclear topology associated with meiotic chromosome pairing were studied in premeiotic cells and spermatocytes I of adult bovine males. To this end, we performed FISH with chromosome, pericentromeric satellite-DNA and telomere-specific probes in combination with immunostaining of synaptonemal complex proteins (SCP3, SCP1) on testis tissue sections. Nuclei of premeiotic cells (spermatogonia) exhibited a scattered telomere distribution while pericentromeres formed a few intranuclear clusters. We observed that the chromosome pairing process in cattle prophase I is preceded by repositioning of centromeres and telomeres to the nuclear periphery during preleptotene. Clustering of chromosome ends (bouquet formation) was observed during the transition from leptonema to zygonema and coincided with pairing of a sub-centromeric marker of bovine chromosomes 7. Dissolution of bouquet topology during zygonema left perinuclear telomeres scattered over the nuclear periphery at pachynema. SCP3 staining in frozen tissue sections revealed the appearance of this axial element protein in intranuclear aggregates during preleptotene, followed by extensive axial element formation during leptotene. Synapsis as revealed by SCP1 staining initiated peripherally at earliest zygotene, at this stage nuclei still contained numerous SCP3 clusters. Our observations reveal prominent non-homologous satellite-DNA associations in spermatogonia and indicate the conservation of topological features of the meiotic chromosome pairing process among mammals. The comparison of telomere dynamics in mouse and cattle prophase I suggests that a larger number of chromosomes prolongs the duration of the bouquet stage.
FISH analysis was used to assign the human ZNF84 gene to chromosome 12q24.33, a region associated with recurrent breakpoints and allelic loss in several human cancers. In this report we show that the ZNF84 coding region is organized in four exons; two are dedicated to encoding the KRAB/FPB-A and KRAB/FPB-B modules, the remaining exons encode the N-terminal amino acids and C-terminal array of zinc finger units, respectively.
The homeobox 11L2 gene (HOX11L2, TLX-3, Rnx) encodes a transcription factor and constitutes together with the HOX11 (TLX) and HOX11L1 (TLX-1, Enx) genes a distinct family of orphan homeobox genes (Kennedy et al., 1991; Roberts et al., 1994; Shirasawa et al., 1997, 2000). In the chick and mouse embryo HOX11L2 is expressed within cranial sensory ganglia, neural crest-derived dorsal root and sympathetic ganglia, as well as in distinct neuronal populations within the hindbrain and the spinal cord (Logan et al., 1998; Shirasawa et al., 2000). Its expression pattern within the developing peripheral and central nervous system suggests that it may be involved in the differentiation of specific neuronal populations as well as in the establishment of neuronal circuitry (Logan et al., 1998). Disruption of the murine homolog of HOX11L2 results in a phenotype resembling that of congenital central hypoventilation syndrome in humans indicating that HOX11L2 is critical for the development of central respiratory structures (Shirasawa et al., 2000). Chromosomal localization of the HOX11L2 gene was carried out as part of its evaluation as a candidate for congenital hypoventilation syndrome.
Cathepsin L (CTSL) is a lysosomal cysteine protease with potent elastase and collagenase activities. Its high activity in the uterine lumen during the period of placental attachment has led to speculation that CTSL may play an important role during embryonic implantation in the pig. Cathepsins have also been implicated in blastocyst implantation in other species like cat, rat and man. We isolated a PAC clone containing the porcine CTSL gene and determined the complete DNA sequence of the gene, which spans about 5.6 kb and consists of eight exons. The CTSL transcript encodes a primary peptide of 334 amino acids sharing 73-78% identity with other mammalian cathepsin L precursor proteins. Based on fluorescence in situ hybridization and radiation hybrid mapping, the porcine CTSL gene was assigned to chromosome 10q11--> q12.
Recently, we cloned a cDNA encoding a novel mouse protein, named A-C1, by differential display between two mouse cell lines, embryonic fibroblast C3H10T1/2 and chondrogenic ATDC5. Mouse A-C1 has homology with a ras-responsive gene, rat Ha-rev107 (Hrasls), and modulates a Ha-ras-mediated signaling pathway. Here, we report a cDNA encoding a human homolog of mouse A-C1. The deduced amino acid sequence of human A-C1 consists of 168 amino acids, and shows 83% identity with that of mouse A-C1. Human A-C1 mRNA was expressed in skeletal muscle, testis, heart, brain, and thyroid in vivo. Moreover, expression of human A-C1 mRNA was detected at a high level in human osteosarcoma-derived U2OS cells in vitro. By FISH analysis the human A-C1 gene (HRASLS) was mapped to human chromosome 3q28→ q29.
Vitamin C (ascorbic acid) is an essential cofactor in several enzymatic reactions (Padh, 1991) and a scavenger of free radicals that protects tissues from oxidative damage (Stadtman and Berlett, 1997). Vitamin C accumulation in tissues follows two pathways. In one, ascorbic acid is oxidized extracellularly to dehydroascorbic acid and transported into the cell by the sodium-independent glucose transporter isoforms GLUT1 and GLUT3 (Rumsey et al., 1997). In the second, ascorbate is directly internalized in a sodium-dependent manner. Recently two sodium-dependent vitamin C transporters, SLC23A1 and SLC23A2 (formerly called SVCT2 and SVCT1, respectively) have been described in rat and man (Daruwala et al., 1999; Tsukaguchi et al., 1999; Wang et al., 2000). Although the two transporters exhibit similar functions and close sequence homology, they differ in tissue distribution. Both in rat and man SLC23A2 had a limited distribution mainly confined to the epithelial system, whereas SLC23A1 is expressed in a host of metabolically active cells and specialized tissues like brain, eye and other organs. Human SLC23A1 and SLC23A2 have been cloned from a kidney cDNA library (Daruwala et al., 1999). The 1953-bp open reading frame of SLC23A1 encodes a 650 amino acid polypeptide. Interestingly SLC23A1 had been previously mapped to human chromosome 20 as an unidentified cDNA sequence, KIAA0238, using a radiation hybrid panel (Nagase et al., 1996). We report here on the chromosomal localization of an EST corresponding to bovine SLC23A1 on BTA Fig. 1. Chromosomal localisation of the bovine SLC23A1 gene. (a) Partial metaphase spread showing the biotinylated SLC23A1 probe on BTA 13q17. (b) Ideogram of BTA13 showing the position of SLC23A1.
Lactotransferrin, a member of the iron binding transferrin family, is present in milk and other biological secretions, as well as in neutrophils. It is involved in host defense against infection and may also play a role in the defense against tumorigenesis (Arnold et al., 1977; Bezault et al., 1994). The LTF gene has previously been assigned to human chromosome 3p21.3 (Kim et al., 1998). Thus mapping of this gene to pig chromosome 13q21 is consistent with ZOO-FISH data showing extensive synteny conservation between these two chromosomes (Gou-
We have cloned and characterised a novel human gene mapping to chromosome 20q11.2. A partial transcript was initially isolated from a human cDNA library transcribed from RNA of the colon carcinoma cell line T-84. In order to determine the full coding sequence of this novel mRNA, we isolated seven cDNA clones from a human cDNA library transcribed from RNA of the acute monocytic leukemia cell line THP1 by colony hybridization. On Northern blot analysis of four human cell lines, the cDNAs isolated hybridize with an abundantly expressed mRNA species of 3.5 kb. A full-length cDNA transcript of this novel mRNA has an open reading frame of 2,796 bp encoding a protein with a calculated molecular weight of 97 kDa. Two repetitive structural consensus motifs are contained within the deduced protein sequence, namely five distinct RNA binding motifs and two proline rich regions. The derived protein sequence also contains putative transmembrane domains. These structural motifs identify this novel protein as a member of an expanding protein family containing RNA binding motifs (RBM). As seen from recently completed sequence of the genomic area encoding this novel mRNA by the Sanger Centre Human Genome Project, the coding region of this gene, RBM12, is intronless.
NFAT5, also known as tonicity enhancer binding protein (TonEBP) or NFATL1, is a new member of the immunologically important NFAT protein family. Despite its obvious relationship to this transcription factor family, NFAT5 shows distinct ways of regulation and function. The complete coding sequence and its alternative splice forms have been described previously. This sequence only refers to less than half of the total mRNA length. High conservation of this gene was shown among man, mouse, and pig. Here we report the cloning of the complete 14-kb cDNA sequence, its genomic organization, and a possible fourth isoform of the corresponding protein. Additionally, we describe the promoter region by CpG-island methylation analysis.
Research Articles| June 28 2001 Isolation and mapping of the pig homolog of the Wiskott-Aldrich Syndrome gene (WAS) Subject Area: Genetics S. Cirera; S. Cirera The Royal Veterinary and Agricultural University, Department of Animal Science and Animal Health, Division of Genetics and Breeding, Frederiksberg (Denmark) Search for other works by this author on: This Site PubMed Google Scholar M. Fredholm M. Fredholm The Royal Veterinary and Agricultural University, Department of Animal Science and Animal Health, Division of Genetics and Breeding, Frederiksberg (Denmark) Search for other works by this author on: This Site PubMed Google Scholar Cytogenetics and Cell Genetics (2001) 92 (3-4): 349–350. https://doi.org/10.1159/000056928 Article history Published Online: June 28 2001 Content Tools Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Tools Icon Tools Get Permissions Cite Icon Cite Search Site Citation S. Cirera, M. Fredholm; Isolation and mapping of the pig homolog of the Wiskott-Aldrich Syndrome gene (WAS). Cytogenetics and Cell Genetics 1 July 2001; 92 (3-4): 349–350. https://doi.org/10.1159/000056928 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsCytogenetic and Genome Research Search Advanced Search This content is only available via PDF. © 2001 S. Karger AG, Basel2001Copyright / Drug Dosage / DisclaimerCopyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements. Article PDF first page preview Close Modal You do not currently have access to this content.
Image restoration approaches, such as digital deconvolution, are becoming widely used for improving the quality of microscopic images. However, no quantification of the gain in resolution of fluorescence images is available. We show that, after iterative constrained deconvolution, fluorescent cosmid signals appear to be 25% smaller, and 1.2-kb fragment signals on combed molecules faithfully display the expected length.
We have determined the organization and chromosome location of the human LANCL1 and mouse Lancl1 genes encoding LANCL1, the lanthionine synthetase component C (LanC)-like protein 1. LANCL1 is related to the bacterial LanC family which is involved in the biosynthesis of antimicrobial peptides. The human and mouse genes span 45 kb and 38 kb, respectively, each comprising ten exons. Within the potential promoter regions, several consensus sequences for ubiquitous and tissue-specific transcription factors are present, reflecting the expression data. The nucleotide sequence of the previously unknown mouse full-length transcript is also reported here. Fluorescence in situ hybridization analyses assigned the LANCL1 gene to human chromosome 2q34 and the Lancl1 gene to mouse chromosome 1, region C2–C5, in accordance with the known homology.
