title: Reassignment of the EPB4.1 gene to 1p36 and assessment of its involvement in neuroblastomas.
abstract: OBJECTIVE
  EPB4.1 has been previously mapped to human chromosome 1p33-p34.2. In contradiction to this chromosomal location, we have mapped EPB4.1-1p36 by using fluorescence in situ hybridization and radiation hybrid mapping. In neuroblastomas, deletions of the telomeric end of chromosome 1 (1p36) are the most common genetic aberration.
  
  
  METHODS
  We investigated whether genetic aberrations of EPB4.1 can be detected in some neuroblastomas by analyzing 72 tumours for EPB4.1 mutation, expression, and alternative splicing pattern. Furthermore, EPB4.1 protein from a neuroblastoma cell line was studied for its subcellular localization.
  
  
  RESULTS
  Sequence changes could be detected in 14 out of 72 neuroblastomas, including missense, silent, and intronic changes. Duplex RT-PCR analysis revealed a subset of 11 tumours expressing significantly low levels of EPB4.1. Significant EPB4.1 sequence changes that were detected included an exon 4 G/A missense mutation (amino acid: V/I) that was shown to be associated with absence of wild-type EPB4.1 expression (3 tumours), an exon 8 G/A missense mutation (V/M) (1 tumour), and an intronic sequence change that was shown to be associated with the presence of an aberrant transcript (1 tumour). Splicing pattern analysis revealed that all EPB4.1 transcripts from tumours exclude exon 3, a splicing pattern for generating the 135 kDa isoform. EPB4.1 cDNA cloned from a neuroblastoma cell line produced a 135-kDa protein with a cytoplasm/membrane localization.
  
  
  CONCLUSIONS
  Out of 72 neuroblastomas we have identified 11 tumours with impaired EPB4.1 expression and 5 tumours with significant sequence changes. We also found that the 135 kDa isoform is the main EPB4.1 product in neuroblastoma. EPB4.1 cDNA from a neuroblastoma cell line produced a 135-kDa protein and displayed a cytoplasm/membrane localization in transfected cells.
authors.affiliation: Molecular Pathogenesis Unit, Surgical Neurology Branch, NINDS/NIH, Building 10/Room 5D32, 9000 Rockville Pike, Bethesda, MD 20892, USA.
authors.forename: S
authors.initials: S
authors.lastname: Huang
authors.forename: U D
authors.initials: UD
authors.lastname: Lichtenauer
authors.forename: S
authors.initials: S
authors.lastname: Pack
authors.forename: C
authors.initials: C
authors.lastname: Wang
authors.forename: A C
authors.initials: AC
authors.lastname: Kim
authors.forename: M
authors.initials: M
authors.lastname: Lutchman
authors.forename: C A
authors.initials: CA
authors.lastname: Koch
authors.forename: J
authors.initials: J
authors.lastname: Torres-Cruz
authors.forename: S C
authors.initials: SC
authors.lastname: Huang
authors.forename: E J
authors.initials: EJ
authors.lastname: Benz
authors.forename: H
authors.initials: H
authors.lastname: Christiansen
authors.forename: B
authors.initials: B
authors.lastname: Dockhorn-Dworniczak
authors.forename: C
authors.initials: C
authors.lastname: Poremba
authors.forename: A O
authors.initials: AO
authors.lastname: Vortmeyer
authors.forename: A H
authors.initials: AH
authors.lastname: Chishti
authors.forename: Z
authors.initials: Z
authors.lastname: Zhuang
mesh_terms: D017398:Alternative Splicing; D001483:Base Sequence; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics*; D003598:Cytoskeletal Proteins; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008566:Membranes / Q000378:metabolism; D009154:Mutation; D009447:Neuroblastoma / Q000235:genetics* / Q000378:metabolism; D009479:Neuropeptides; D020033:Protein Isoforms / Q000235:genetics; D014407:Tumor Cells, Cultured
chemical_list: D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D004273:DNA, Neoplasm; D008565:Membrane Proteins; D009479:Neuropeptides; D020033:Protein Isoforms; C028841:erythrocyte membrane band 4.1 protein; C120557:erythrocyte membrane protein band 4.1-like 1