Life Sciences Commons^™

Fanconi Anemia Protein Fancd2 Inhibits Trf1 Polyadp-Ribosylation Through Tankyrase1-Dependent Manner, Alex Lyakhovich, Maria Ramirez, Andres Castella, Amanda Simons, Jeffrey Parvin, Jordi Suralles

Amanda Simons

Background: Fanconi anemia (FA) is a rare autosomal recessive syndrome characterized by developmental abnormalities, progressive bone marrow failure, and predisposition to cancer. The key FA protein FANCD2 crosstalks with members of DNA damage and repair pathways that also play a role at telomeres. Therefore, we investigated whether FANCD2 has a similar involvement at telomeres. Results: We reveal that FANCD2 may perform a novel function separate to the FANCD2/BRCA pathway. This function includes FANCD2 interaction with one of the telomere components, the PARP family member tankyrase-1. Moreover, FANCD2 inhibits tankyrase-1 activity in vitro. In turn, FANCD2 deficiency increases the polyADPribosylation of …

Identification Of Genetic Risk Associated With Prostate Cancer Using Ancestry Informative Markers, Bradford Wilson

Bradford Wilson

The Molecular Basis Of Drug Resistance Against Hepatitis C Virus Ns3/4a Protease Inhibitors, Keith Romano, Akbar Ali, Cihan Aydin, Djade Soumana, Aysegul Ozen, Laura Deveau, Casey Silver, Hong Cao, Alicia Newton, Christos Petropoulos, Wei Huang, Celia Schiffer

Celia A. Schiffer

Hepatitis C virus (HCV) infects over 170 million people worldwide and is the leading cause of chronic liver diseases, including cirrhosis, liver failure, and liver cancer. Available antiviral therapies cause severe side effects and are effective only for a subset of patients, though treatment outcomes have recently been improved by the combination therapy now including boceprevir and telaprevir, which inhibit the viral NS3/4A protease. Despite extensive efforts to develop more potent next-generation protease inhibitors, however, the long-term efficacy of this drug class is challenged by the rapid emergence of resistance. Single-site mutations at protease residues R155, A156 and D168 confer …

Long-Term Correction Of Very Long-Chain Acyl-Coa Dehydrogenase Deficiency In Mice Using Aav9 Gene Therapy, Allison Keeler, Thomas Conlon, Glenn Walter, Huadong Zeng, Scott Shaffer, Fu Dungtao, Kirsten Erger, Travis Cossette, Qiushi Tang, Christian Mueller, Terence Flotte

Christian Mueller

Very long-chain acyl-coA dehydrogenase (VLCAD) is the rate-limiting step in mitochondrial fatty acid oxidation. VLCAD-deficient mice and patients clinical symptoms stem from not only an energy deficiency but also long-chain metabolite accumulations. VLCAD-deficient mice were treated systemically with 1 x 10(12) vector genomes of recombinant adeno-associated virus 9 (rAAV9)-VLCAD. Biochemical correction was observed in vector-treated mice beginning 2 weeks postinjection, as characterized by a significant drop in long-chain fatty acyl accumulates in whole blood after an overnight fast. Changes persisted through the termination point around 20 weeks postinjection. Magnetic resonance spectroscopy (MRS) and tandem mass spectrometry (MS/MS) revealed normalization of …

Use Of Highly Polymorphic Dna Probes For Genotypic Analysis Following Bone Marrow Transplantation, Robert Knowlton, Valerie Brown, Jeffrey Braman, David Barker, James Schumm, Christine Murray, Tak Takvorian, Jerome Ritz, Helen Donis-Keller

Helen Donis-Keller

The use of DNA markers known as restriction fragment length polymorphisms is a sensitive and informative method of distinguishing patient and allogeneic donor cells after bone marrow transplantation. To apply the test, it is necessary in each case to find DNA probes that display patient-specific and donor-specific bands in Southern transfer hybridization. We have isolated a set of 12 cloned DNAs from highly polymorphic loci by which siblings can usually be distinguished. With just four of these probes, we can expect to distinguish the genotypes of the recipient and a sibling donor in more than 99% of cases (except between …

Mutations In The Ret Proto-Oncogene Are Associated With Men 2a And Fmtc, Helen Donis-Keller, Shenshen Dou, David Chi, Katrin Carlson, Koi Toshima, Terry Lairmore, James Howe, Jeffrey Moley, Paul Goodfellow, Samuel Wells

Helen Donis-Keller

Multiple endocrine neoplasia type 2A (MEN 2A) and familial medullary thyroid carcinoma (FMTC) are dominantly inherited conditions which predispose to the development of endocrine neoplasia. Evidence is presented that sequence changes within the coding region of the RET proto-oncogene, a putative transmembrane tyrosine kinase, may be responsible for the development of neoplasia in these inherited disorders. Single strand conformational variants (SSCVs) in exons 7 and 8 of the RET proto-oncogene were identified in eight MEN 2A and four FMTC families. The variants were observed only in the DNA of individuals who were either affected or who had inherited the MEN2A …

Single Missense Mutation In The Tyrosine Kinase Catalytic Domain Of The Ret Protooncogene Is Associated With Multiple Endocrine Neoplasia Type 2b., Katrin Carlson, Shenshen Dou, David Chi, N. Scavarda, Koi Toshima, C. Jackson, Samuel Wells, Paul Goodfellow, Helen Donis-Keller

Helen Donis-Keller

Multiple endocrine neoplasia type 2B (MEN 2B) is a human cancer syndrome characterized by medullary thyroid carcinoma (MTC), pheochromocytomas, mucosal neuromas, ganglioneuromas of the intestinal tract, and skeletal and ophthalmic abnormalities. It appears both as an inherited disorder and as de novo disease. Sequence analysis of germ-line DNA from MEN 2B patients revealed the existence of the same point mutation in the RET protooncogene in 34 unrelated individuals. This sequence difference was not observed in 93 unaffected individuals, including the normal parents of 14 de novo MEN 2B patients. The mutation (ATG-->ACG) results in the replacement of methionine with …

A Genetic Linkage Map Of The Human Genome, Helen Donis-Keller, Philip Green, Cynthia Helmes, Samuel Cartinhour, Barbara Weiffenbach, Karen Stephens, Tim Keith, Donald Bowden, Douglas Smith, Eric Lander, David Botstein, Gita Akots, Kenneth Rediker, Thomas Gravius, Valerie Brown, Marcia Rising, Carol Parker, Jody Powers, Diane Watt, Erick Kauffman, Angela Bricker, Pamela Phipps, Hans Muller-Kahle, Thomas Fulton, Siu Ng, James Schumm, Jeffrey Braman, Robert Knowlton, David Barker, Steven Crooks, Steven Lincoln, Mark Daly, Jeff Abrahamson

Helen Donis-Keller

We report the construction of a linkage map of the human genome, based on the pattern of inheritance of 403 polymorphic loci, including 393 RFLPs, in a panel of DNAs from 21 three-generation families. By a combination of mathematical linkage analysis and physical localization of selected clones, it was possible to arrange these loci into linkage groups representing 23 human chromosomes. We estimate that the linkage map is detectably linked to at least 95% of the DNA in the human genome.

Genetic Analysis Of Eight Loci Tightly Linked To Neurofibromatosis 1, Karen Stephens, Philip Green, V. Riccardi, Siu Ng, Marcia Rising, David Barker, J. Darby, K. Falls, F. Collins, H. Willard, Helen Donis-Keller

Helen Donis-Keller

The genetic locus for neurofibromatosis 1 (NF1) has recently been mapped to the pericentromeric region of chromosome 17. We have genotyped eight previously identified RFLP probes on 50 NF1 families to determine the placement of the NF1 locus relative to the RFLP loci. Thirty-eight recombination events in the pericentromeric region were identified, eight involving crossovers between NF1 and loci on either chromosomal arm. Multipoint linkage analysis resulted in the unique placement of six loci at odds greater than 100:1 in the order of pter-A10-41-EW301-NF1-EW207-CRI-L581-CRI-L946 -qter. Owing to insufficient crossovers, three loci--D17Z1, EW206, and EW203--could not be uniquely localized. In this …

A Polymorphic Dna Marker Linked To Cystic Fibrosis Is Located On Chromosome 7, Robert Knowlton, Odile Cohen-Haguenauer, Nguyen Van Cong, Jean Frézal, Valerie Brown, David Barker, Jeffrey Braman, James Schumm, Lap-Chee Tsui, Manuel Buchwald, Helen Donis-Keller

Helen Donis-Keller

Although cystic fibrosis (CF) is among the most common inherited diseases in Caucasian populations, the basic biochemical defect is not yet known. CF is inherited as an autosomal recessive trait apparently due to mutations in a single gene, whence the efforts made to identify the genetic locus responsible by linkage studies. Two markers have recently been identified that are genetically linked to CF: one is a genetic variation in serum level of activity of the enzyme paraoxonase, and the other is a restriction fragment length polymorphism (RFLP) identified with a randomly isolated DNA probe. We report here that the genetic …

Chromosomal Bar Codes Produced By Multicolor Fluorescence In Situ Hybridization With Multiple Yac Clones And Whole Chromosome Painting Probes, Christoph Lengauer, Michael Speicher, Susanne Popp, Anna Jauch, Masafumi Taniwaki, Ramaiah Nagaraja, Harold Riethman, Helen Donis-Keller, Michele D'Urso, David Schelssinger, Thomas Cremer

Helen Donis-Keller

Colored chromosome staining patterns, termed chromosomal ‘bar codes’ (CBCs), were obtained on human chromosomes by fluorescence in situhybridization (FISH) with pools of Alu-PCR products from YAC dones containing human DNA inserts ranging from 100 kbp to 1 Mbp. In contrast to conventional G- or R-bands, the chromosomal position, extent, Individual color and relative signal intensity of each ‘bar’ could be modified depending on probe selection and labeling procedures. Alu-PCR amplification products were generated from 31 YAC clones which mapped to 37 different chromosome bands. For multiple color FISH, Alu-PCR amplification products from various clones were either biotinylated or labeled …

Predictive Dna Testing And Prophylactic Thyroidectomy In Patients At Risk For Multiple Endocrine Neoplasia Type 2a, Samuel Wells, David Chi, Koi Toshima, Louis Dehner, Cheryl Coffin, S. Dowton, Jennifer Ivanovich, Mary Debenedettl, William Dilley, Jeffrey Moley, Jeffrey Norton, Helen Donis-Keller

Helen Donis-Keller

Background: Missense germ-line mutations in the RET protooncogene are associated with multiple endocrine neoplasia type 2A (MEN 2A). Detection of these mutant alleles in kindred members predicts disease inheritance and provides the basis for preventative thyroidectomy. Methods: A polymerase chain reaction (PCR)-based genetic test for the 19 known RET mutations was designed to study 132 members of 7 kindreds with MEN 2A. Haplotypes also were constructed using genetic markers flanking the MEN 2A locus. Plasma calcitonin (CT) concentrations were determined before and after provocative testing. Results: Direct DNA testing and haplotype analysis showed that 21 of 58 kindred members at …

Gene Transfer In The Lung Using Recombinant Adeno-Associated Virus, Alisha Gruntman, Christian Mueller, Terence Flotte, Guangping Gao

Christian Mueller

Adeno-associated virus (AAV) is a small replication-deficient DNA virus belonging to the Parvovirinae family. It has a single-stranded ∼4.7-kb genome. Recombinant AAV (rAAV) is created by replacing the viral rep and cap genes with the transgene of interest along with promoter and polyadenylation sequences. The short viral inverted terminal repeats must remain intact for replication and packaging in production, as well as vector genome processing and persistence in the transduction process. The AAV capsid (serotype) determines the tissue tropism of the rAAV vector. In this unit we will discuss serotype selection for lung targeting along with the factors effecting efficient …

A Unified View Of Base Excision Repair, Karen Almeida, Robert Sobol

Karen H Almeida

Base excision repair (BER) proteins act upon a significantly broad spectrum of DNA lesions that result from endogenous and exogenous sources. Multiple sub-pathways of BER (short-path or longpatch) and newly designated DNA repair pathways (e.g., SSBR and NIR) that utilize BER proteins complicate any comprehensive understanding of BER and its role in genome maintenance, chemotherapeutic response, neurodegeneration, cancer or aging. Herein, we propose a unified model of BER, comprised of three functional processes: Lesion Recognition/Strand Scission, Gap Tailoring and DNA Synthesis/Ligation, each represented by one or more multiprotein complexes and coordinated via the XRCC1/DNA Ligase III and PARP1 scaffold proteins. …

Identification Of Sonic Hedgehog As A Candidate Gene Responsible For Holoprosencephaly, Helen Donis-Keller, E Belloni, M Muenke, E Roessler, G Traverse, J Siegel-Bartelt, A Frumkin, H Mitchell, C Helms, A Hing, H Heng, B Kroop, D Martindale, J Rommens, L Tsui, S Scherer

Helen Donis-Keller

Holoprosencephaly (HPE) is a genetically and phenotypically heterogenous disorder involving the development of forebrain and midface, with an incidence of 1:16,000 live born and 1:250 induced abortions. This disorder is associated with several distinct facies and phenotypic variability: in the most extreme cases, anophthalmia or cyclopia is evident along with a congenital absence of the mature nose. The less severe form features facial dysmorphia characterized by ocular hypertelorism, defects of the upper lip and/or nose, and absence of the olfactory nerves or corpus callosum. Several intermediate phenotypes involving both the brain and face have been described. One of the gene …

7q11.23 Deletions In Williams Syndrome Arise As A Consequence Of Unequal Meiotic Crossover, Helen Donis-Keller, Zsolt Urbán, C Helms, G Fekete, Katalin Csiszár, D Bonnet, A Munnich, C Boyd

Helen Donis-Keller

Williams syndrome (WS) (Williams et al. 1961; Beurenet al. 1962) is a multisystem disorder characterized by mental retardation, a specific neurobehavioral profile,characteristic facies, infantile hypercalcemia, cardiovascular abnormalities, progressive joint limitation, hernias, and soft skin. Recent studies have shown that hemizygosity at the elastin (ELN) gene locus on chromosome7q is associated with WS (Ewart et al. 1993). Furthermore, two FISH studies using cosmid recombinants containing the 5' or the 3' end of the ELN gene revealed deletion of the entire ELN gene in 90%-96% of classical WS cases (Lowery et al. 1995; Nickerson et al. 1995). However, the size of the …

Comparative Genomic Hybridization Of Human Malignant Gliomas Reveals Multiple Amplification Sites And Nonrandom Chromosomal Gains And Losses, Helen Donis-Keller, Evelin Schròck, Gundula Thiel, Tanka Lozanova, Stanislas Du Manoir, Marie-Christine Meffert, Anna Jauch, Michael Speicher, Peter Nürnberg, Siegfried Vogel, Werner Janisch, Thomas Ried, Regine Witkowski, Thomas Cremer

Helen Donis-Keller

Nine human malignant gliomas (2 astrocytomas grade III and 7 glioblastomas) were analyzed using comparative genomic hybridization (CGH). In addition to the amplification of the EGFR gene at 7p12 in 4 of 9 cases, six new amplification sites were mapped to 1q32, 4q12, 7q21.1, 7q21.2-3, 12p, and 22q12. Nonrandom chromosomal gains and losses were identified with overrepresentation of chromosome 7 and underrepresentation of chromosome 10 as the most frequent events (1 of 2 astrocytomas, 7 of 7 glioblastomas). Gain of a part or the whole chromosome 19 and losses of chromosome bands 9pter-23 and 22q13 were detected each in five …

Recombinant Mapping Of The Familial Hyperinsulinism Gene To An 0.8 Cm Region On Chromosome 11p15.1 And Demonstration Of A Founder Effect In Ashkenazi Jews, Helen Donis-Keller, Benjamin Glaser, Ken Chiu, Li Liu, Roberto Anker, Ann Nestorowicz, Nancy Cox, Heddy Landau, Kalser Nurit, Paul Thornton, Charles Stanley, Erol Cerasl, Lester Baker, M. Alan Permutt

Helen Donis-Keller

A gene for autosomal recessive familial hyperinsulinism (HI) (OMIM: 256450), a neonatal metabolic disease characterized by inappropriate insulin secretion in the presence of severe hypoglycemia, was recently mapped to a 6.6 cM interval between the markers D11S926 and D11S928 onchromosome 11p in 15 families (1). In the current study we evaluated six additional families and five new markers, and further localized the gene between D11S419 and D11S1310. Using genotype data from CEPH Version 7 and data generated from this study, this region was estimated to be 0.8 cM in length. Significant linkage disequilibrium between markers and the HIgene was observed …

Sequence-Ready Contig For The 1.4-Cm Ductal Carcinoma In Situ Loss Of Heterozygosity Region On Chromosome 8p22–P23, Helen Donis-Keller, Jen Wang, Diane Radford, Matthew Holt, C Helms, A Goate, W Brandt, M Parik, Nancy Phillips, K Deschryver, M Schuh, Keri Fair, Jon Ritter, P Marshall

Helen Donis-Keller

We report the construction of an ∼1.7-Mb sequence-ready YAC/BAC clone contig of 8p22–p23. This chromosomal region has been associated with frequent loss of heterozygosity (LOH) in breast, ovarian, prostate, head and neck, and liver cancer. We first constructed a meiotic linkage map for 8p to resolve previously reported conflicting map orders from the literature. The target region containing a putative tumor suppressor gene was defined by allelotyping 65 cases of sporadic ductal carcinoma in situ with 18 polymorphic markers from 8p. The minimal region of loss encompassed the interval between D8S520 and D8S261, and one tumor had loss of D8S550 …

Isolation, Characterization, And Chromosomal Mapping Of The Human Nkx6.1 Gene (Nkx6a), A New Pancreatic Islet Homeobox Gene, Helen Donis-Keller, Hiroshi Inoue, Abraham Rudnick, Michael German, Rosalie Veile, M. Alan Permutt

Helen Donis-Keller

Nkx6.1 (gene symbol NKX6A), a new member of the NK homeobox gene family, was recently identified in rodent pancreatic islet β-cell lines. The pattern of expression suggested that this gene product might be important for control of islet development and/or regulation of insulin biosynthesis. We now report cloning of human NKX6A, characterization of its genomic structure, and its chromosomal localization. The predicted protein of human NKX6A contained 367 amino acids and had 97% identity to the hamster protein. The highly conserved NK decapeptide and homeodomain regions were identical between human and hamster, suggesting functional importance of these domains. The coding …

Identification Of A Human Lmx1 (Lmx1.1)-Related Gene, Lmx1.2: Tissue-Specific Expression And Linkage Mapping On Chromosome 9, Helen Donis-Keller, Christopher Iannotti, Hiroshi Inoue, Ernesto Bernal-Mizrachi, Minoru Aoki, Li Liu, Michael German, M. Alan Permutt

Helen Donis-Keller

LMX1 is a LIM-homeodomain (LIM-HD)-containing protein expressed selectively in insulin-producing β-cell lines, and it it has been shown to activate insulin gene transcription. The human LMX1 gene was mapped by fluorescencein situhybridization to chromosome region 1q22–q23, yet Churchet al.(1994,Nat. Genet.6: 98–105) identified two exon-trapping products from human chromosome 9 that were highly homologous to hamster LMX1. In the current study, we demonstrate tissue-specific expression of an LMX1 (now known as LMX1.1)-related gene, named LMX1.2. The chicken C-LMX1 gene, recently cloned using the hamster LMX1.1 sequence and shown to specify dorsal cell fate during vertebrate …

Supravalvular Aortic Stenosis: A Splice Site Mutation Within The Elastin Gene Results In Reduced Expression Of Two Aberrantly Spliced Transcripts, Helen Donis-Keller, Zsolt Urbán, Virginia Michels, Stephen Thibodeau, Katalin Csiszár, C Boyd

Helen Donis-Keller

We have screened the elastin gene for mutations responsible for supravalvular aortic stenosis (SVAS) in two large, independently collected families with isolated (nonsyndromic) SVAS. By single-strand conformation polymorphism and heteroduplex analysis, we have identified a C to G transversion within the acceptor splice site of exon 16 in SVAS patients from both families. This mutation segregates in both families with high penetrance of SVAS, and all affected individuals carry the mutation. Haplotype analysis by using closely linked polymorphisms, including a previously unreported BfaI restriction fragment length polymorphism within the 3’-UTR of the elastin gene, indicates that the mutations found …

High Levels Of Loss At The 17p Telomere Suggest The Close Proximity Of A Tumour Suppressor, Helen Donis-Keller, G White, M Stack, M Santibáñez-Koref, D Liscia, T Venesio, Jen Wang, C Helms, D Betticher, H Altermatt, P Hoban, J Heighway

Helen Donis-Keller

High levels of loss of distal markers on 17p13.3 in breast cancer suggested the presence within the region of at least one tumour-suppressor gene. Here we describe the derivation of two biallelic polymorphisms from the 17p telomeric yeast artificial chromosome (YAC) TYAC98. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiplex PCR analysis demonstrated that the high level of allelic imbalance observed in breast tumours represented loss of constitutional heterozygosity (LOH) and that this LOH extended to the telomere. Lung carcinoma (but not Wilms' tumour)-derived DNA again revealed a high level of loss of subtelomeric 17p sequences. Telomeric microsatellite polymorphisms …

Genetic Fine Mapping Of The Gene For Nonsyndromic Congenital Retinal Nonattachment., Helen Donis-Keller, N Ghiasvand, T Fleming, C Helms, Aledavood Avisa

Helen Donis-Keller

Autosomal recessive nonsyndromic congenital retinal nonattachment (NCRNA) comprises congenital insensitivity to light, massive retrolental mass, shallow anterior chamber, microphthalmia, and nystagmus in otherwise normal individuals. Polymerase chain reaction-based linkage analyses of polymorphic microsatellite markers in the 10q21 region on DNA samples from 106 individuals provide evidence that the NCRNA locus is within an interval of ∼0.6–1.5 cM, flanked by the markers D10S522 and D10S1418. Haplotype analysis demonstrated a unique founder haplotype shared by 100% of the NCRNA chromosomes. These results indicate a founder effect and the strong possibility of a single mutation as the cause of the disease in the …

Mapping Human Telomere Regions With Yac And P1 Clones: Chromosome-Specific Markers For 27 Telomeres Including 149 Stss And 24 Polymorphisms For 14 Proterminal Regions, Helen Donis-Keller, A Vocero-Akbani, C Helms, Jen Wang, F Sanjurjo, J Korte-Sarfaty, Rosalie Veile, L Liu, A Jauch, A Burgess, A Hing, Matthew Holt, S Ramachandra, A Whelan, R Anker, L Ahrent, M Chen, M Gavin, K Iannantuoni, S Morton, Sunil Pandit, C Read, Todd Steinbrueck, Christopher Warlick, D Smoller

Helen Donis-Keller

A YAC library enriched for telomere clones was constructed and screened for the humantelomere-specific repeat sequence (TTAGGG). Altogether 196 TYAC library clones were studied: 189 new TYAC clones were isolated, 149 STSs were developed for 132 different TY-ACs, and 39P1 clones were identified using 19 STSs from 16 of the TYACs. A combination of mappingmethods including fluorescence in situ hybridization, somatic cell hybrid panels, clamped homogeneous electric fields, meiotic linkage, and BLASTN sequence analysis was utilized to characterize the resource. Forty-five of the TYACs map to 31 specific telomere regions. Twenty-four linkage markers were developed and mapped within 14 proterminal …

Subregional Localization Of 21 Chromosome 7-Specific Expressed Sequence Tags (Ests) By Fish Using Newly Identified Yacs And P1s, Helen Donis-Keller, S Morton, Rosalie Veile, Cynthia Helms, Martin Lee, Wen-Lin Kuo, Joe Gray

Helen Donis-Keller

Twenty-one putative chromosome 7-derived expressed sequence tags (ESTs) identified 33 yeast artificial chromosomes (YACs) or P1 clones, which were then used as reagents for physical mapping. FISH mapping established that the ESTs contained within these clones were distributed throughout chromosome 7, with all major cytogenetic bands represented, except 7p13–p15, 7p11, 7q31.2, and 7q35. Each EST sequence identified at least one other sequence in publicly available databases (using search tools such as BLASTN, basic local alignment search tool), and many of the ESTs identified cDNAs and several genomic DNA sequences. However, 7 ESTs did not identify highly significant matches (P< 1 …

The Finding Of A Somatic Deletion In Ret Exon 15 Clarified The Sporadic Nature Of A Medullary Thyroid Carcinoma Suspected To Be Familial, Helen Donis-Keller, J Oriola, I Halperin, F Rivera-Fillat

Helen Donis-Keller

Medullary thyroid carcinoma (MTC) occurs both sporadically and in the autosomal dominantly inherited multiple endocrine neoplasia (MEN) type 2 syndromes. The distinction between both is important for future clinical management. We report a family initially described as a familial MTC by pentagastrin stimulation test and clinical outcome, in which we found a 12 bp deletion within the catalytic domain of the protooncogene RET in the index case tumor alone. Linkage study suggests that it is a sporadic MTC. Therefore, in view of these results, in kindred with just one MTC case, borderline pentagastrin test values must be carefully assessed. In …

The Multiple Endocrine Neoplasia Type 2b Point Mutation Alters Long-Term Regulation And Enhances The Transforming Capacity Of The Epidermal Growth Factor Receptor, Helen Donis-Keller, Sunil Pandit, Takeo Iwamoto, John Tomich, Linda Pike

Helen Donis-Keller

The RET proto-oncogene encodes a member of the receptor tyrosine kinase family. Multiple endocrine neoplasia type 2B (MEN 2B) is caused by the mutation of a conserved methionine to a threonine in the catalytic domain of the RET kinase. When the MEN 2B point mutation was introduced into the epidermal growth factor (EGF) receptor (M857T EGFR), the intrinsic tyrosine kinase activity of the mutant receptor was similar to that of wild-type EGF receptor and remained ligand-dependent. However, the mutant receptor showed an enhanced transforming capacitycompared to the wild-type receptor as judged by its ability to mediate the growth of NIH …

Index, Comprehensive Microsatellite, And Unified Linkage Maps Of Human Chromosome 14 With Cytogenetic Tie Points And A Telomere Microsatellite Marker, Helen Donis-Keller, Sunil Pandit, Jen Wang, Rosalie Veile, Santosh Mishra, Christopher Warlick

Helen Donis-Keller

Three sets of linkage maps (index, comprehensive microsatellite, and unified) have been constructed for human chromosome 14 based on genotypes from the CEPH reference pedigrees. The index maps consist of 18 microsatellite markers, with heterozygosities of at least 68% and intermarker spacing no greater than 11 cM. The sex-average comprehensive microsatellite map is 125 cM in length and includes 115 markers with 54 loci uniquely placed with odds for marker order of at least 1000:1. The sex-average index map length is 121 cM, and the female- and male-specificmaps are 143 and 101 cM, respectively. A unified map was also constructed …

The Ceph Consortium Linkage Map Of Human Chromosome 11, M Litt, P Kramer, E. Kort, P Fain, S Cox, D Root, R White, J Weissenbach, Helen Donis-Keller, R Gatti, J Weber, Y Nakamura, C Julier, K Hayashi, N Spurr, M Dean, J Mandel, K Kidd, T Kruse, A Retief, A Bale, T Meo, G Vergnaud, S Warren, H Willard

Helen Donis-Keller

The CEPH consortium framework map of chromosome 11 is presented. The map was generated from CEPH family DNAs with 181 probe/enzyme combinations contributed by 20 laboratories. Seventy-seven of the loci are defined by microsatellite polymorphisms that can be typed by the PCR. A total of 42 loci have been placed on the map with likelihood support of at least 1000:1. The female, male, and sex-average maps extend for 179.6, 110.8, and 145.3 cM, respectively. The largest interval on the sex-average map is less than 11 cM, and the average distance between uniquely placed loci is 4 cM. The genotypic data …