DOCSLIB.ORG

Somatic Deletion Mapping on Chromosome 10 and Sequence Analysis of PTEN/MMAC1 Point to the 10Q25-26 Region As the Primary Target in Low-Grade and High-Grade Gliomas

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Somatic Deletion Mapping on Chromosome 10 and Sequence Analysis of PTEN/MMAC1 Point to the 10Q25-26 Region As the Primary Target in Low-Grade and High-Grade Gliomas Oncogene (1998) 16, 3331 ± 3335 1998 Stockton Press All rights reserved 0950 ± 9232/98 $12.00 http://www.stockton-press.co.uk/onc SHORT REPORT Somatic deletion mapping on chromosome 10 and sequence analysis of PTEN/MMAC1 point to the 10q25-26 region as the primary target in low-grade and high-grade gliomas Daniel Maier1, Zuwen Zhang1, Elisabeth Taylor1, Marie-France Hamou2, Otmar Gratzl1, Erwin G Van Meir2, Rodney J Scott1 and Adrian Merlo1 1Molecular Neuro-Oncology, Neurosurgery and Department of Research, University Hospital, Schanzenstr.46, CH-4031 Basel; 2Laboratory of Tumor Biology and Genetics, Department of Neurosurgery, University Hospital, CH-1011 Lausanne, Switzerland The 10q25-26 region between the dinucleotide markers dicult to delineate speci®c regions of loss, 10q25-26 D10S587 and D10S216 is deleted in glioblastomas and, has consistently been suspected to harbor a tumor as we have recently shown, in low-grade oligodendro- suppressor gene (Fults et al., 1993; Rasheed et al., gliomas. We further re®ned somatic mapping on 10q23- 1995; Albarosa et al., 1996). Recently, PTEN/MMAC1 tel and simultaneously assessed the role of the candidate on 10q23.3 has been proposed as a candidate tumor tumor suppressor gene PTEN/MMAC1 in glial neo- suppressor gene for gliomas and other neoplasms, plasms by sequence analysis of eight low-grade and 24 clearly de®ning a second locus on 10q (Li et al., 1997; high-grade gliomas. These tumors were selected for Steck et al., 1997). This gene has also been shown to be partial or complete loss of chromosome 10 based on involved in two rare inherited disorders, the Cowden deletion mapping with increased microsatellite marker (Liaw et al., 1997) and the Bannayan-Zonana density at 10q23-tel. Three out of eight (38%) low-grade syndrome (Marsh et al., 1997). PTEN/MMAC1 and 3/24 (13%) high-grade gliomas exclusively target encodes a protein tyrosine phosphatase, shares 10q25-26. We did not ®nd a tumor only targeting extensive homology to the cytoskeletal proteins tensin 10q23.3, and most tumors (23/32, 72%) showed large and auxilin, and has been shown to act either as deletions on 10q including both regions. The sequence positive or negative regulator during signal transduc- analysis of PTEN/MMAC1 revealed nucleotide altera- tion, cell cycle progression, and cellular transformation tions in 1/8 (12.5%) low-grade gliomas in a tumor with (Tonks et al., 1996), and is regulated by transforming LOH at 10q21-qtel and in 5/21 (24%) high-grade growth factor b (Li and Sun, 1997). gliomas displaying LOH that always included 10q23- The more telomeric region on 10q25-26 between the 26. Our re®ned mapping data point to the 10q25-26 dinucleotide markers D10S587 and D10S216 is region as the primary target on 10q, an area that also involved in glioblastomas (Rasheed et al., 1995; harbors the DMBT1 candidate tumor suppressor gene. Albarosa et al., 1996), and in low-grade oligoden- The fact that we ®nd hemizygous deletions at 10q25-qtel drogliomas, too, as we have recently shown (Maier et in low-grade astrocytomas and oligodendrogliomas ± two al., 1997). The involvement of this very region in histologically distinct entities of gliomas ± suggests the earlier tumor stages has also been described in existence of a putative suppressor gene involved early in endometrial carcinomas (Nagase et al., 1997). We glial tumorigenesis. further re®ned somatic mapping on 10q in another seven low-grade gliomas (®ve astrocytomas and two Keywords: deletion mapping; chromosome 10q23.3 and mixed oligoastrocytomas). From the total number 10q25-26; PTEN/MMAC1 (n=86) of low-grade (n=14) and high-grade (n=72) gliomas which have been used for somatic mapping on chromosome 10, we selected eight low-grade (®ve astrocytomas, one mixed oligoastrocytoma, and two Loss of heterozygosity (LOH) on chromosome 10 has oligodendrogliomas) and 24 high-grade gliomas, which frequently been found in a variety of human cancers, either displayed LOH at the PTEN/MMAC1 locus such as endometrial (Peier et al., 1995), prostate (n=23), selectively targeted 10q25-26 (n=6) or were of (Gray et al., 1995), bladder (Cappellen et al., 1997), low-grade type without LOH on chromosome 10 renal cell carcinoma (Morita et al., 1991), Non- (n=3), to assess the role of PTEN/MMAC1 in glial Hodgkin lymphomas (Speaks et al., 1992), malignant tumorigenesis by analysing the genomic sequence of meningiomas (Rempel et al., 1993), melanomas (Herbst this new candidate suppressor gene on 10q23.3. These et al., 1994) and gliomas (Fults et al., 1990; Fujimoto tumors were selected for partial and complete LOH, et al., 1989; Rasheed et al., 1992; James et al., 1988; since the biallelic inactivation of a tumor suppressor Steck et al., 1995). Eighty to ninety per cent of gene typically involves an intragenic change (nucleotide malignant gliomas display large deletions on chromo- substitution, small insertion, or microdeletion) within some 10. Although somatic mapping has proven one allele, combined with inactivation of the other allele through the loss of a large chromosomal region (Knudson, 1993). Sequence analysis of PTEN/MMAC1 Correspondence: A Merlo revealed nucleotide alterations in 1/8 (12.5%) low- Received 22 October 1997; revised 8 January 1998; accepted 8 grade gliomas and in 5/21 (24%) high-grade gliomas. January 1998 None of the 9/32 (28%) tumors that did not display Two targets on chromosome 10q23-26 in gliomas DMaieret al 3332 LOH at the PTEN/MMAC1 locus showed sequence oligodendrogliomas and ®ve astrocytomas). DNA from alterations of this gene. All mutated tumors displayed tumor specimens and lymphocytes was analysed for LOH at the PTEN/MMAC1 locus. These ®ndings loss of heterozygosity (LOH) by ampli®cation of 31 di- suggest that PTEN/MMAC1 plays a role in glioma and tetranucleotide repeat-containing sequences using tumorigenesis. However, we ®nd partial somatic PCR and the conditions described (Merlo et al., 1994). deletions to be more frequent on 10q25-26 in low- Primers for microsatellite markers were obtained from grade and high-grade gliomas, suggesting that this Research Genetics (Huntsville, AL). Primer sequences region is the primary target in glioma tumorigenesis on for PTENCA, a highly informative dinucleotide repeat chromosome 10q. marker at the 5' end of PTEN/MMAC1, were kindly Tumor tissue and peripheral blood lymphocytes provided by P Cairns and D Sidransky (Wang et al., were obtained from 32 patients diagnosed with 1997). For microsatellite analysis, one primer was primary CNS tumors and were classi®ed according to labeled with T4-polynucleotide kinase (New England the World Health Organization: 24 high-grade astro- Biolabs) and g-32P-ATP. Fifty nanograms of genomic cytomas grade IV (glioblastoma multiforme) and eight DNA (normal and tumor) were subjected to 30 cycles low-grade gliomas grade II (one oligoastrocytoma, two of PCR ampli®cation with annealing temperatures that Table 1 Mutations and polymorphisms of PTEN/MMAC1 in primary gliomas Tumor type Case no. Codon changea Mutation Exon/Intron Predicted alterationb All GE31 CAT183→CGT183 missense 3 H61R GBM BS7 ATG594→D[ATG]592 ± 594 frameshift 6 delM198 GBM BS10 T1G intron 4 splicing variant GBM BS11 CGA699→TGA699 nonsense 7 R233Opal BS11 del T intron 3 polymorphism GBM BS54 CGC519→CAC519 missense 6 R173H GBM LS971 TAT264→D[AT]263 ± 264 frameshift 5 Y88fs a Mutated nucleotides are in boldface type. b Positions refer to the deduced PTEN protein sequence; fs, frameshift b ACGT a NNTT q 21.1 q 25.2 D10S676 D10S221 q 23.3 q 25.3 PTEN/MMAC1 locus c ACGT PTENCA D10S587 q 24.2 q 26.1 D10S254 D10S214 Figure 1 LOH analysis on chromosome 10q and mutations of PTEN/MMAC1 in primary gliomas (a) The deletion breakpoint in this low-grade astrocytoma (GE74) lies telomeric to the PTEN/MMAC1 locus (10q23.3) between the markers D10S254 (10q24.2) and D10S221 (10q25.2). Arrowheads denote loss of alleles consistent with LOH. N, normal; T, tumor. (b) Mutation in glioblastoma (GBM7, middle lane; GBM2, left lane; GBM8, right lane). Sequence of nucleotides 571 ± 603 of exon 6 of PTEN/MMAC1 in the antisense orientation. Arrow indicates an in frame 3 bp deletion of nucleotides 592 ± 594. (c) Mutation in a low-grade astrocytoma (GE31). Sequence of nucleotides 173 ± 193 of genomic DNA from blood (left) and primary tumor (right) from the same patient using a sense primer. Arrow indicates a transversion from A to G at nucleotide 182 Two targets on chromosome 10q23-26 in gliomas DMaieret al 3333 ranged from 50 ± 628C. Products were separated using Li et al. in their original reports. The fact that we 7% polyacrylamide-formamide gel electrophoresis detected a mutation in a low-grade astrocytoma followed by autoradiography. For informative cases, (GE31) suggests that this candidate tumor suppressor allelic loss was scored if the radiographic signal of one gene might occasionally also contribute to earlier allele was at least 50% less in tumor DNA as stages of tumorigenesis. Since the mutation rate as compared to the corresponding normal allele. assessed by direct analysis of the genomic sequence of For mutation screening of the candidate tumor PTEN/MMAC1 in primary gliomas was found to be suppressor gene PTEN/MMAC1, 50 ng of genomic much lower than LOH at the PTEN/MMAC1 locus, DNA from primary tumors and blood lymphocytes we searched for alternative mechanisms of gene was ampli®ed by PCR. Cycle sequencing was inactivation, that could be missed by direct sequence performed directly on PCR products using a nested analysis. For example, homozygous deletions are sequencing primer end-labeled with g32P-ATP and the technically dicult to exclude in primary tumor AmpliCycleTM Sequencing Kit (Perkin Elmer, Branch- specimens. It has been shown that an apparently burg, NJ).
Load more

Recommended publications
  • Identification of a Region of Homozygous Deletion on 8P22–23.1 in Medulloblastoma
    Oncogene (2002) 21, 1461 ± 1468 ã 2002 Nature Publishing Group All rights reserved 0950 ± 9232/02 $25.00 www.nature.com/onc Identi®cation of a region of homozygous deletion on 8p22 ± 23.1 in medulloblastoma Xiao-lu Yin1, Jesse Chung-sean Pang1 and Ho-keung Ng*,1 1Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China To identify critical tumor suppressor loci that are radiotherapy and chemotherapy, have greatly improved associated with the development of medulloblastoma, the outcomes of patients. In the past 30 years, the 5- we performed a comprehensive genome-wide allelotype year survival rate has increased from 10 to about 50%. analysis in a series of 12 medulloblastomas. Non-random However, long-term survival in children with advanced allelic imbalances were identi®ed on chromosomes 7q disease is only about 30% (Giangaspero et al., 2000; (58.3%), 8p (66.7%), 16q (58.3%), 17p (58.3%) and Heideman et al., 1997). Further enhancement of 17q (66.7%). Comparative genomic hybridization analy- survival will rely on a better understanding of the sis con®rmed that allelic imbalances on 8p, 16q and 17p biology of this malignant disease to improve current were due to loss of genetic materials. Finer deletion treatments or to develop novel therapy. mapping in an expanded series of 23 medulloblastomas Non-random loss of genetic material from chromo- localized the common deletion region on 8p to an interval somal loci is a common feature in the development of of 18.14 cM on 8p22 ± 23.2.
    [Show full text]
  • Salmonella Typhimurium
    A REFINED MAP OF THE hisG GENE OF SALMONELLA TYPHIMURIUM I. HOPPE, H. M. JOHNSTON, D. BIEK AND J. R. ROTH Department of Biology, University of Utah, Salt Lake City, Utah 84112 Manuscript received October 2, 1978 ABSTRACT The hisG gene is the most operator-proximal structural gene of the histi­ dine operon; it encodes the feedback-inhibitable first enzyme of the biosyn­ thetic pathway. Previously, hisG mutants were mapped into seven intervals defined by the available deletion mutations having endpoints in the hisG gene. The map has been refined using over 60 new deletion mutants. The new map divides the gene into 40 deletion intervals, which average approximately 30 base pairs in length. The map has been used to analyze the distribution of insertion sites for the transposable element TntO and has permitted con­ clusions on the distribution of duplication endpoints. The map promises to be useful in analysis of his regulation and, more particularly, in the determina­ tion of the possible role of the hisG enzyme in this mechanism. THE hisG gene is the most operator-proximal gene of the histidine operon (BRENNER and AMES 1971; HARTMAN, HARTMAN and STAHL 1971). It en­ codes the structure of the first enzyme in histidine biosynthesis, PR-ATP synthe­ tase (BRENNER and AMES 1971). This enzyme has been purified (MARTIN 1963; WHITFIELD 1971; BLASI, ALOJ and GOLDBERGER 1971; VOLL, APELLA and MAR­ TIN 1967; PARSONS and KOSHLAND 1974) and intensively analysed because of its sensitivity to feedback inhibition (MARTIN 1963; WHITFIELD 1971; MORTON and PARSONS 1977) and its complex subunit structure (PARSONS and KOSHLAND 1974a,b).
    [Show full text]
  • Deletion Mapping of Chromosome 4 in Head and Neck Squamous Cell Carcinoma
    Oncogene (1997) 14, 369 ± 373 1997 Stockton Press All rights reserved 0950 ± 9232/97 $12.00 Deletion mapping of chromosome 4 in head and neck squamous cell carcinoma Mark A Pershouse1,3, Adel K El-Naggar2, Kenneth Hurr2, Huai Lin1,3, WK Alfred Yung1,3 and Peter A Steck1,3 Departments of 1Neuro-Oncology and 2Pathology and 3The Brain Tumor Center, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA Genomic deletions involving chromosome 4 have recently Cytogenetic studies have identi®ed recurring, but been implicated in several human cancers. To identify widely varied alterations of chromosomes 1, 3, 4, 5, 7, and characterize genetic events associated with the 8, 9, 11, 14, 15 and 17 (Jin et al., 1993; Sreekantaiah et development of head and neck squamous cell carinoma al., 1994). Although several molecular studies have (HNSCC), a ®ne mapping of allelic losses associated shown that mutation of p53 and ampli®cation of with chromosome 4 was performed on DNA isolated epidermal growth factor receptor are relatively from 27 matched primary tumor specimens and normal common events. However, the exact genes that are tissues. Loss of heterozygosity (LOH) of at least one targeted in the majority of the observed chromosomal chromosome 4 polymorphic allele was seen in the alterations are unknown (Brachman et al., 1992; majority of tumors (92%). Allelic deletions were con®ned Grandis et al., 1993; Shin et al., 1994). Recently, to short arm loci in four tumors and to the long arm loci several groups have performed allelotyping studies on in 12 tumors, suggesting the presence of two regions of HNSCC specimens to further de®ne regions of common deletion.
    [Show full text]
  • Isolation of DNA Markers in the Direction of the Huntington Disease Gene from the G8 Locus
    Am. J. Hum. Genet. 42:335-344, 1988 Isolation of DNA Markers in the Direction of the Huntington Disease Gene from the G8 Locus Barbara Smith,* Douglas Skarecky,* Ulla Bengtsson,* R. Ellen Magenis,t Nancy Carpenter,: and John J. Wasmuth* *Department of Biological Chemistry, California College of Medicine, University of California, Irvine; tDepartment of Medical Genetics, Crippled Children's Division, Oregon Health Sciences University, Portland; and tH. Allen Chapman Research Institute of Medical Genetics, Children's Medical Center, Tulsa Summary To facilitate identification of additional DNA markers near and on opposite sides of the Huntington dis- ease (HD) gene, we developed a panel of somatic-cell hybrids that allows accurate subregional mapping of DNA fragments in the distal portion of 4p. By means of the hybrid-cell mapping panel and a library of DNA fragments enriched for sequences from the terminal one-third of the short arm of chromosome 4, 105 DNA fragments were mapped to six different physical regions within 4pl5-4pter. Four polymorphic DNA fragments of particular interest were identified, at least three of which are distal to the HD-linked D4S10 (G8) locus, a region of 4p previously devoid of DNA markers. Since the HD gene has also recently been shown to be distal to G8, these newly identified DNA markers are in the direction of the HD gene from G8, and one or more of them may be on the opposite side of HD from G8. Introduction diagnosed, they are past the reproductive age, and Huntington disease (HD) is an inherited, autosomal each child of an affected individual is at 50% risk for dominant neuropsychiatric disorder.
    [Show full text]
  • Cyclin D1: Mechanism and Consequence of Androgen Receptor Co-Repressor Activity in Prostatic Adenocarcinoma
    UNIVERSITY OF CINCINNATI Date: April 1st, 2004 I, __________________Christin E. Petre_________________________, hereby submit this work as part of the requirements for the degree of: Doctor of Philosophy in: Cell and Molecular Biology It is entitled: Cyclin D1: Mechanism and Consequence of Androgen Receptor Co-repressor Activity in Prostatic Adenocarcinoma This work and its defense approved by: Chair: Karen E. Knudsen, Ph.D. Sohaib Khan, Ph.D. Kenji Fukasawa, Ph.D. Alvaro Puga, Ph.D. Linda Parysek, Ph.D. J. Alan Diehl, Ph.D. Robert Hennigan, Ph.D. CYCLIN D1: MECHANISM AND CONSEQUENCE OF ANDROGEN RECEPTOR CO-REPRESSOR ACTIVITY IN PROSTATIC ADENOCARCINOMA A dissertation submitted to the Division of Research and Advanced Studies of the University of Cincinnati in partial fulfillment of the requirements for the degree of DOCTORATE OF PHILOSOPHY (Ph.D.) in the Department of Cell Biology, Neurobiology, and Anatomy of the College of Medicine 2004 by Christin E. Petre B.A., Miami University, 2000 Committee Chair: Karen E. Knudsen, Ph.D. ABSTRACT It is increasingly evident that androgen receptor (AR) regulation plays a critical role in the development and progression of prostate cancer. We show that induction of cyclin D1 occurs upon ligand stimulation in androgen dependent prostatic adenocarcinoma (LNCaP) cells. Such induction results in the formation of active cyclin dependent kinase (CDK)-cyclin D1 complexes, phosphorylation of the rentinoblastoma tumor suppressor protein, and concomitant cell cycle progression. In addition, we find that cyclin D1 harbors a second cell cycle independent function responsible for restraining AR transactivation. We illustrate that cyclin D1 co-repressor activity is extremely potent, inhibiting receptor transactivation independently of cellular background, promoter context, co-activator over expression, ligand/non-ligand activators, and cancer predisposing AR mutations/polymorphisms.
    [Show full text]
  • Mapping Genes for X-Linked Disorders
    r6's'êf1 [l MAPPING GENES FOR X.LINKED DISORDERS by Ági Kyra Gedeon B.Sc.(Hons.) This thesis is submitted as the complete requirement for the degree of Doctor of Philosophy within the Department of Genetics, Faculty of Science, at The University of Adelaide, South Australia. September 1996 Declaration This thesis contains no material that has been accepted for the award of any other degree or diploma in any University or other tertiary institution, nor to the best of the candidate's knowledge any material previously published or written by another person, except where due reference is made in the text. Throughout this volume any reference to 'the candidate' signifies the author Agi Kyra Gedeon. The work described in this thesis has been performed by the candidate or under the candidate's supervision, except where duly acknowledged in the text. NAME: A.q4 ..K.,,..6.ç.ù.çp-M.'...." couRSE: F¿. Þ.,..ßçi.c.r.c*) v in the university Libraries' being available I give consent to this copy of my thesis, when deposited for photocoPying and loan. 2..ï..: !9.:1.ú SIGNATURE: ... DArE: Table of Contents Declaration Table of Contents 1l Abbreviations iii Acknowledgments v Summary vii Note regarding Publications ix CHAPTER 1 : Gene Mapping: Review of the literature and Objectives 1 CHAPTER 2 : Materials and Methods JJ CHAPTER 3 : Evolution of the Background Map of the X Chromosome 62 CHAPTER 4 : Non-specific X-linked Mental Retardation (MRX) 92 CHAPTER 5 : Syndromal X-linked Mental Retardation (XLMR) r46 CHAPTER 6 : Other X-linked Disorders 167 CHAPTER
    [Show full text]
  • Deletion Mapping of Four Loci Defined by N-Ethyl-N-Nitrosourea-Induced Postimplantation-Lethal Mutations Within the Pid-Hb
    Copyright 0 1993 by the Genetics Society of America Deletion Mapping of Four Loci Defined by N-Ethyl-N-Nitrosourea-Induced Postimplantation-Lethal Mutations Within thepid-Hbb Region of Mouse Chromosome 7 Eugene M. Rinchik, Donald A. Carpenter and Carol L. Long Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831-8077 Manuscript received May 12, 1993 Accepted for publication August 21, 1993 ABSTRACT As part of a long-term effort to refine the physical and functional maps of the Fes-Hbb region of mouse chromosome 7,four loci [1(7)1Rn, 1(7)2Rn,1(7)3Rn, 1(7)4Rn] defined by N-ethyl-N-nitrosourea (ENU)-induced, prenatally lethal mutationswere mapped by means of trans complementation crosses to mice carrying lethal deletionsof the mouse chromosome-7 albino(c) locus. Each locuswas assigned to a defined subregionof the deletionmap at the distal endof the Fes-Hbb interval. Of particular use for this mapping were preimplantation-lethal deletions having distal breakpoints localized between pad and Omp. Hemizygosity or homozygosity for each of the ENU-induced lethalswas found to arrest development after uterine implantation; the specifictime of postimplantation death varied, and depended on both the mutation itself and on whether it was hemizygous or homozygous. Based on their map positions outsideof and distal to deletions that cause death at preimplantation stages, these ENU-induced mutations identify loci, necessary for postimplantation development, that could not have been discovered by phenotypic analyses of mice homozygous for any albino deletion. The mapping of these loci to specific genetic intervals definedby deletion breakpoints suggestsa number of positional-cloning strategies for the molecular isolation of these genes.
    [Show full text]
  • The Importance of Mutation, Then and Now: Studies with Yeast Cytochrome C
    2 F. Sherman / Mutation Research 589 (2005) 1–16 Mutation Research 589 (2005) 1–16 Contents 1. My first encounter with yeast research . .................................................. 2 The importance of mutation, then and now: 2. Identification of CYC1 ............................................................... 3 Mutation Research 589 (2005) 1–16 ☆ 3. Importance of cytochrome c ........................................................... 4 studies with yeast cytochrome c www.elsevier.com/locate/reviewsmr Community address: www.elsevier.com/locate/mutres Fred Sherman * Reflections in Mutation Research 4. CYC1 encodes iso-1-cytochrome c ....................................................... 4 5. Isolation and characterization of cyc1 mutants. .............................................. 5 Department ofThe Biochemistry importance and Biophysics, of University mutation, of Rochester then Medical andSchool, now:Box 712, Rochester, studies NY 14642, USA Accepted 26 July 2004 with yeast cytochrome c$ 6. Generating altered genes and proteins, then and now .......................................... 6 Available online 23 September 2004 7. Deducing DNA sequences from protein sequences. .......................................... 8 Fred Sherman* 8. ‘‘Site-directed’’ mutagenesis in early times . .............................................. 10 Department of Biochemistry and Biophysics, University of Rochester Medical School, Box 712, Rochester, NY 14642, USA 9. Cloning CYC1 and transcriptional regulation. .............................................
    [Show full text]
  • The Art and Design of Genetic Screens: Mouse
    Nature Reviews Genetics | AOP, published online 10 June 2005; doi:10.1038/nrg1636 REVIEWS THE ART AND DESIGN OF GENETIC SCREENS: MOUSE Benjamin T. Kile and Douglas J. Hilton Abstract | Humans are mammals, not bacteria or plants, yeast or nematodes, insects or fish. Mice are also mammals, but unlike gorilla and goat, fox and ferret, giraffe and jackal, they are suited perfectly to the laboratory environment and genetic experimentation. In this review, we will summarize the tools, tricks and techniques for executing forward genetic screens in the mouse and argue that this approach is now accessible to most biologists, rather than being the sole domain of large national facilities and specialized genetics laboratories. MOUSE FANCY Not surprisingly, given its experimental advantages steps of the screening process: mutagenesis, screen A collection of enthusiasts BOX 1, the mouse has been by our side as we have design and execution, and to its dénouement: the interested in mice with unusual walked down the road of biological self-discovery. identification of the causative genetic change. traits. Mouse mutants have been a source of fascination for millennia1. A century ago, the mouse was there with Mutagenesis Cuénot to provide the first confirmation that Mendel’s Historically, mouse genetics has relied on sponta- newly rediscovered laws applied to animals as well neous mutation for introducing genetic variety into as plants2. Since then, the mouse has become the pre- populations being screened. A steady stream of mouse eminent model for human physiology and disease. A mutants was identified in wild populations, both few years ago, the sequence of the mouse genome was among the collections of the MOUSE FANCY and in the completed, providing a detailed, if furry, reflection of colonies maintained at the Jackson Laboratory.
    [Show full text]
  • High-Resolution Deletion Mapping of 15Q13.2-Q21.1 in Transitional Cell Carcinoma of the Bladder
    [CANCER RESEARCH 63, 7657–7662, November 15, 2003] High-Resolution Deletion Mapping of 15q13.2-q21.1 in Transitional Cell Carcinoma of the Bladder Rachael Natrajan, Jari Louhelainen, Sarah Williams, Jonathan Laye, and Margaret A. Knowles Division of Cancer Medicine Research, Cancer Research UK Clinical Centre, St. James’s University Hospital, Leeds, United Kingdom ABSTRACT 3p, 6p, 8p, 8q, 10p, 10q, 11q, 12q, 17q, and 20q. In contrast, in papillary superficial tumors, few alterations have been found at high Deletions found in several types of human tumor, including carcinomas frequency, with the exception of deletions of chromosome 9 found in of the colorectum, breast, and lung, suggest the presence of a potential ϳ50% of cases (3) and mutation of the fibroblast growth factor tumor suppressor gene(s) on chromosome 15. Common regions of deletion in these tumors are at 15q15 and 15q21. Here, we have analyzed loss of receptor 3 gene (FGFR3) found in 70% (4). Because bladder cancer heterozygosity (LOH) on chromosome 15 to ascertain its potential involve- is a disease of middle and old age, it is predicted that additional ment in the development and progression of transitional cell carcinoma heritable alterations contribute to the development of superficial blad- (TCC) of the bladder. A panel of 26 polymorphic markers, spanning der tumors, and these remain to be identified. 15q12–15q22, were used to map regions of LOH in 51 TCCs. LOH was We have previously performed an allelotype analysis of TCC (5), found for at least one marker in the region 15q14–15q15.3 in 20 of 51 and there have been several comprehensive CGH and cytogenetic (39%) tumors.
    [Show full text]
  • Current Status of Human Chromosome 14 D Kamnasaran,Dwcox
    81 REVIEW ARTICLE J Med Genet: first published as 10.1136/jmg.39.2.81 on 1 February 2002. Downloaded from Current status of human chromosome 14 D Kamnasaran,DWCox ............................................................................................................................. J Med Genet 2002;39:81–90 Over the past three decades, extensive genetic, GENETIC, PHYSICAL, TRANSCRIPT AND physical, transcript, and sequence maps have assisted SEQUENCE MAPPING RESOURCES FOR CHROMOSOME 14 in the mapping of over 30 genetic diseases and in the Genetic maps identification of over 550 genes on human chromosome Genetic maps of the human genome have been 14. Additional genetic disorders were assigned to constructed to increase the informativeness and density of genetic markers. Refined chromosome chromosome 14 by studying either constitutional or 14 specific linkage maps were made by integrat- acquired chromosome aberrations of affected subjects. ing markers from different genetic maps and by Studies of benign and malignant tumours by karyotype reassessing the genotype data. The progress in mapping was summarised annually at the Gene analyses and by allelotyping with a panel of Mapping Meetings and two Chromosome 14 polymorphic genetic markers have further suggested the Workshops were held.10 11 Early chromosome 14 presence of several tumour suppressor loci on genetic maps had very few blood serotype, RFLP, and VNTR genetic markers, and were biased with chromosome 14. The search for disease genes on the highest density of markers in the distal
    [Show full text]
  • A DELETION MAP of Cycl MUTANTS and ITS CORRESPONDENCE to MUTATIONALLY ALTERED ISO-1-CYTOCHROMES C of YEAST 1975). Even
    A DELETION MAP OF cycl MUTANTS AND ITS CORRESPONDENCE TO MUTATIONALLY ALTERED ISO-1-CYTOCHROMES c OF YEAST FRED SHERMAN, MARY JACKSON, SUSAN W. LIEBMAN, ANN MARIE SCHWEINGRUBERI AND JOHN W. STEWART Department of Radiation Biology and Biophysics University of Rochester School of Medicine and Dentistry, Rochester, New York 14642 Manuscript received March 14, 1975 ABSTRACT Mutants arising spontaneously from sporulated cultures of certain strains of yeast, Saccharomyces cereuisiae, contained deletions of the CYCl gene which controls the primary structure of iso-1-cytochrome c. At least 60 different kinds of deletions were uncovered among the 104 deletions examined and these ranged in length from those encompassing only two adjacent point mutants to those encompassing at least the entire CYCl gene. X-ray-induced recombina- tion rates of crosses involving these deletions and cycl point mutants resulted in the assignment of 211 point mutants to 47 mutational sites and made it possible to unambiguously order 40 of these 47 sites. Except for one mutant, cycl-15, there was a strict colinear relationship between the deletion map and the positions of 13 sites that were previously determined by amino acid altera- tions in iso-1-cytochromes c from intragenic revertants. ECENTLY we described 210 single-site mutants of the CYCl gene which de- R termines the primary structure of iso-l -cytochrome c in Saccharomyces cereuisiae (SHERMANet al. 1974). A consistent genetic map could not be con- structed from X-ray-induced recombination rates of over 2000 allelic pairs of these cycl mutants, even though this procedure is most often used for fine-struc- ture mapping of yeast mutants.
    [Show full text]