TY - JOUR
T1 - cDNA sequence and genomic structure of EVI2B, a gene lying within an intron of the neurofibromatosis type 1 gene
AU - Cawthon, Richard M.
AU - Andersen, Lone B.
AU - Buchberg, Arthur M.
AU - Xu, Gangfeng
AU - O'Connell, Peter
AU - Viskochil, David
AU - Weiss, Robert B.
AU - Wallace, Margaret R.
AU - Marchuk, Douglas A.
AU - Culver, Melanie
AU - Stevens, Jeffrey
AU - Jenkins, Nancy A.
AU - Copeland, Neal G.
AU - Collins, Francis S.
AU - White, Ray
N1 - Funding Information:
Lymphoblastoid cell lines from normal and NFl individuals, established by transformation of peripheral blood mononuclear cells with Epstein-Barr virus, and somatic cell hybrid lines containing various portions of chromosome 17 were maintained in culture as described by O’Connell et al. (1990). Skin fibroblasts were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, and passaged every 5-7 days with a trypsin/EDTA solution. Mononuclear cells were prepared from whole peripheral blood by the LeucoPREP procedure according to the manufacturer’s instructions (Becton Dickinson and Company, Lincoln Park, NJ). Bone marrow aspirates (performed at, University Hospital, University of Utah Medical Center, with informed consent of the patients and Institutional Review Board approval) drawn into tubes with heparin were spun at 2000 rpm for 10 min in a tabletop centrifuge; the pellets were then used for preparation of total RNA (see below). Other tissues removed from patients for therapeutic reasons or at autopsy were immediately placed in liquid nitrogen and stored at -135°C until used for RNA preparation. Some tissue specimens were obtained from the National Neurological Research Bank, VAMC Wadsworth (Los Angeles, CA) which is sponsored by NINCDS/NIMH, NMSS, HD Foundation, TS Association, and the Veterans Administration.
Funding Information:
We thank V. M. Riccardi and D. Fults for providing NFl tissues: M. Robertson for the fluorescent sequencing of EVf2B cDNAs on the Applied Biosystems Inc. Model 370A DNA sequencer and K. Murphy for assistance with the manual sequencing; and S. Noth-wehr for analyzing EVIZH with the SIGSEQ programs. R. Foltz prepared the tigures. This study was supported in part by the National Cancer Institute, Department of Health and Human Services, under Contract Ntll-CO-74101 with ABL and in part hy NIH Grant NS23410 to F.S.C. L.B.A. is a recipient of a fellowship from the llniversity of Copenhagen, Denmark; D.V. and M.R.W. are supported by the National Neurofihromatosis Foundation; R.M.C. is an Associate, P.O’C. a Senior Associate, F.S.C. an Associate Investigator, and R.W. an Investigator in the Howard Hughes Medical Institute.
PY - 1991/3
Y1 - 1991/3
N2 - The gene responsible for neurofibromatosis type 1 (NF1), one of the more common inherited human disorders, was identified recently, and segments of it were cloned. Two translocation breakpoints that interrupt the NF1 gene in NF1 patients flank a 60-kb segment of DNA that contains the EVI2A locus (previously reported as the EVI2 locus), the human homolog of a mouse gene, Evi-2A, implicated in retrovirus-induced murine myeloid tumors. EVI2A lies within an intron of the NF1 gene and is transcribed from telomere toward centromere, opposite to the direction of transcription of the NF1 gene. Here we describe a second locus, EVI2B, also located between the two NF1 translocation breakpoints. Full-length cDNAs from the EVI2B locus detect a 2.1-kb transcript in bone marrow, peripheral blood mononuclear cells, and fibroblasts. Sequencing studies predict an EVI2B protein of 448 amino acids that is proline-rich and contains an N-terminal signal peptide, an extracellular domain with four potential glycosylation sites, a single hydrophobic transmembrane domain, and a cytoplasmic hydrophilic domain. At the level of genomic DNA the EVI2B locus lies within the same intron of the NF1 gene as EVI2A and contains a 57-bp 5′ exon that is noncoding, an 8-kb intron, and a 2078-bp 3′ exon that includes the entire open reading frame. EVI2B is transcribed in the same direction as EVI2A; its 5′ exon lies only 4 kb downstream from the 3′ exon of the EVI2A locus. In the mouse the 5′ exon of the homologous gene, Evi-2B, lies approximately 2.8 kb from the 3′ end of Evi-2A, in the midst of a cluster of viral integration sites identified in retrovirus-induced myeloid tumors; thus, Evi-2B may function as an oncogene in these tumors.
AB - The gene responsible for neurofibromatosis type 1 (NF1), one of the more common inherited human disorders, was identified recently, and segments of it were cloned. Two translocation breakpoints that interrupt the NF1 gene in NF1 patients flank a 60-kb segment of DNA that contains the EVI2A locus (previously reported as the EVI2 locus), the human homolog of a mouse gene, Evi-2A, implicated in retrovirus-induced murine myeloid tumors. EVI2A lies within an intron of the NF1 gene and is transcribed from telomere toward centromere, opposite to the direction of transcription of the NF1 gene. Here we describe a second locus, EVI2B, also located between the two NF1 translocation breakpoints. Full-length cDNAs from the EVI2B locus detect a 2.1-kb transcript in bone marrow, peripheral blood mononuclear cells, and fibroblasts. Sequencing studies predict an EVI2B protein of 448 amino acids that is proline-rich and contains an N-terminal signal peptide, an extracellular domain with four potential glycosylation sites, a single hydrophobic transmembrane domain, and a cytoplasmic hydrophilic domain. At the level of genomic DNA the EVI2B locus lies within the same intron of the NF1 gene as EVI2A and contains a 57-bp 5′ exon that is noncoding, an 8-kb intron, and a 2078-bp 3′ exon that includes the entire open reading frame. EVI2B is transcribed in the same direction as EVI2A; its 5′ exon lies only 4 kb downstream from the 3′ exon of the EVI2A locus. In the mouse the 5′ exon of the homologous gene, Evi-2B, lies approximately 2.8 kb from the 3′ end of Evi-2A, in the midst of a cluster of viral integration sites identified in retrovirus-induced myeloid tumors; thus, Evi-2B may function as an oncogene in these tumors.
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U2 - 10.1016/0888-7543(91)90410-G
DO - 10.1016/0888-7543(91)90410-G
M3 - Article
C2 - 1903357
AN - SCOPUS:0025978395
VL - 9
SP - 446
EP - 460
JO - Genomics
JF - Genomics
SN - 0888-7543
IS - 3
ER -