Coamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG.

2.50
Hdl Handle:
http://hdl.handle.net/10541/95595
Title:
Coamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG.
Authors:
Heighway, Jim; Betticher, Daniel C; Hoban, Paul R; Altermatt, H J; Cowen, Rachel L
Abstract:
Analysis of a region of DNA, coamplified in tumors with KRAS2, resulted in the identification of the human homologue of the mouse KRAG gene. The gene was widely expressed in a range of cell lines, tumors, and normal tissue and demonstrated a high degree of alternate splicing. A human KRAG cDNA sequence, with a structure similar to that encoded by the amplified gene in mouse Y1 adrenal carcinoma cells, was isolated by RT-PCR. The predicted amino acid similarity between the two sequences was 91%, and hydrophobicity plots suggested a structure closely resembling that of transmembrane 4 superfamily members. Identification of a PCR-based restriction fragment length polymorphism confirmed biallelic expression of KRAG but suggested allele-specific splicing differences in tumors. Northern analysis of mRNA derived from a range of tissues suggested high level expression in muscle and confirmed alternate splicing. To facilitate the analysis of exon junctions, a YAC clone encoding the genomic sequence was identified. This allowed the localization of KRAG to human chromosome 12p11.2. Isolation of one end of this nonchimeric clone demonstrated a perfect match with a 247-bp sequence within the 3' untranslated region of the type 2 1,4, 5-inositol triphosphate receptor gene. Multiplex PCR confirmed the inclusion of both genes in the KRAS2 amplicon in human malignancy, suggesting that either may contribute to the malignant phenotype.
Affiliation:
CRC Department of Cancer Genetics, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Wilmslow Road, Manchester, M20 9BX, United Kingdom.
Citation:
Coamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG. 1996, 35 (1):207-14 Genomics
Journal:
Genomics
Issue Date:
1-Jul-1996
URI:
http://hdl.handle.net/10541/95595
DOI:
10.1006/geno.1996.0340
PubMed ID:
8661122
Type:
Article
Language:
en
ISSN:
0888-7543
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorHeighway, Jimen
dc.contributor.authorBetticher, Daniel Cen
dc.contributor.authorHoban, Paul Ren
dc.contributor.authorAltermatt, H Jen
dc.contributor.authorCowen, Rachel Len
dc.date.accessioned2010-04-06T08:50:54Z-
dc.date.available2010-04-06T08:50:54Z-
dc.date.issued1996-07-01-
dc.identifier.citationCoamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG. 1996, 35 (1):207-14 Genomicsen
dc.identifier.issn0888-7543-
dc.identifier.pmid8661122-
dc.identifier.doi10.1006/geno.1996.0340-
dc.identifier.urihttp://hdl.handle.net/10541/95595-
dc.description.abstractAnalysis of a region of DNA, coamplified in tumors with KRAS2, resulted in the identification of the human homologue of the mouse KRAG gene. The gene was widely expressed in a range of cell lines, tumors, and normal tissue and demonstrated a high degree of alternate splicing. A human KRAG cDNA sequence, with a structure similar to that encoded by the amplified gene in mouse Y1 adrenal carcinoma cells, was isolated by RT-PCR. The predicted amino acid similarity between the two sequences was 91%, and hydrophobicity plots suggested a structure closely resembling that of transmembrane 4 superfamily members. Identification of a PCR-based restriction fragment length polymorphism confirmed biallelic expression of KRAG but suggested allele-specific splicing differences in tumors. Northern analysis of mRNA derived from a range of tissues suggested high level expression in muscle and confirmed alternate splicing. To facilitate the analysis of exon junctions, a YAC clone encoding the genomic sequence was identified. This allowed the localization of KRAG to human chromosome 12p11.2. Isolation of one end of this nonchimeric clone demonstrated a perfect match with a 247-bp sequence within the 3' untranslated region of the type 2 1,4, 5-inositol triphosphate receptor gene. Multiplex PCR confirmed the inclusion of both genes in the KRAS2 amplicon in human malignancy, suggesting that either may contribute to the malignant phenotype.en
dc.language.isoenen
dc.subjectCanceren
dc.subjectCancer DNAen
dc.subjectCancer Proteinsen
dc.subjectCancer RNAen
dc.subjectLung Canceren
dc.subjectCultured Tumour Cellsen
dc.subject.meshAlleles-
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshCalcium Channels-
dc.subject.meshCarcinoma, Non-Small-Cell Lung-
dc.subject.meshCarcinoma, Squamous Cell-
dc.subject.meshCarrier Proteins-
dc.subject.meshCell Transformation, Neoplastic-
dc.subject.meshChromosomes, Human, Pair 12-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA, Complementary-
dc.subject.meshDNA, Neoplasm-
dc.subject.meshGene Amplification-
dc.subject.meshGene Expression Regulation, Neoplastic-
dc.subject.meshGenes, ras-
dc.subject.meshHumans-
dc.subject.meshInositol 1,4,5-Trisphosphate Receptors-
dc.subject.meshLung Neoplasms-
dc.subject.meshMembrane Proteins-
dc.subject.meshMice-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshNeoplasm Proteins-
dc.subject.meshNeoplasms-
dc.subject.meshRNA Splicing-
dc.subject.meshRNA, Messenger-
dc.subject.meshRNA, Neoplasm-
dc.subject.meshReceptors, Cytoplasmic and Nuclear-
dc.subject.meshSequence Alignment-
dc.subject.meshSequence Homology, Amino Acid-
dc.subject.meshSpecies Specificity-
dc.subject.meshTumor Cells, Cultured-
dc.titleCoamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Cancer Genetics, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Wilmslow Road, Manchester, M20 9BX, United Kingdom.en
dc.identifier.journalGenomicsen

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