title: Panhandle polymerase chain reaction amplifies MLL genomic translocation breakpoint involving unknown partner gene.
abstract: We used a new approach called panhandle polymerase chain reaction (PCR) to clone an MLL genomic translocation breakpoint in a case of acute lymphoblastic leukemia of infancy in which karyotype analysis was technically unsuccessful and did not show the translocation partner. Panhandle PCR amplified known MLL sequence 5' of the breakpoint and 3' sequence from the unknown partner gene from a DNA template with an intrastrand loop schematically shaped like a pan with a handle. The 7-kb panhandle PCR product contained the translocation breakpoint in MLL intron 8. The partner DNA included unique nonrepetitive sequences, Alu and mammalian apparent LTR-retrotransposon (MaLR) repetitive sequences, and a region of homology to expressed sequence tags. MaLR sequences have not been found before near leukemia-associated translocation breakpoints. The nonrepetitive sequences were not homologous to known partner genes of MLL. Screening of somatic cell hybrid and radiation hybrid lines by PCR and fluorescence in situ hybridization analysis of normal metaphase chromosomes mapped the partner DNA to chromosome band 4q21. Reverse transcriptase-PCR identified an MLL-AF-4 chimeric mRNA, indicating that panhandle PCR identified a fusion of MLL with a previously uncharacterized AF-4 intronic sequence. Panhandle PCR facilitates cloning translocation breakpoints and identifying unknown partner genes.
authors.affiliation: Division of Oncology, Leonard and Madlyn Abramson Pediatric Research Center, Children's Hospital of Philadelphia, PA 19104-4318, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Felix
authors.forename: C S
authors.initials: CS
authors.lastname: Kim
authors.forename: M D
authors.initials: MD
authors.lastname: Megonigal
authors.forename: D J
authors.initials: DJ
authors.lastname: Slater
authors.forename: D H
authors.initials: DH
authors.lastname: Jones
authors.forename: N B
authors.initials: NB
authors.lastname: Spinner
authors.forename: T
authors.initials: T
authors.lastname: Stump
authors.forename: M R
authors.initials: MR
authors.lastname: Hosler
authors.forename: P C
authors.initials: PC
authors.lastname: Nowell
authors.forename: B J
authors.initials: BJ
authors.lastname: Lange
authors.forename: E F
authors.initials: EF
authors.lastname: Rappaport
mesh_terms: D001483:Base Sequence; D002894:Chromosomes, Human, Pair 4; D004268:DNA-Binding Proteins / Q000235:genetics*; D005260:Female; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D007223:Infant; D007438:Introns; D008969:Molecular Sequence Data; D051788:Myeloid-Lymphoid Leukemia Protein; D016133:Polymerase Chain Reaction / Q000379:methods*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D011519:Proto-Oncogenes; D014157:Transcription Factors; D014178:Translocation, Genetic
chemical_list: D004268:DNA-Binding Proteins; C497422:KMT2A protein, human; D014157:Transcription Factors; D051788:Myeloid-Lymphoid Leukemia Protein; D011495:Histone-Lysine N-Methyltransferase