A 6-Mb yeast artificial chromosome contig and long-range physical map encompassing the region on chromosome 12q15 frequently rearranged in a variety of benign solid tumors.
Cytogenetic analysis of a variety of benign solid tumors, among which uterine leiomyoma, lipoma, pleomorphic salivary gland adenoma, and pulmonary chondroid hamartoma, has indicated that these tumors often display chromosome breakpoints in region q13-q15 of chromosome 12. In previous studies, we have reported that these breakpoints map between locus D12S8 and the CHOP gene, the latter of which has been shown to be consistently rearranged in myxoid liposarcomas with t(12;16)(q13;p11). Here, we report directional chromosome walking studies starting from D12S8 and resulting in the construction of a YAC contig of about 6 Mb. This YAC contig, whose orientation on chromosome 12 was determined by double-color fluorescence in situ hybridization (FISH) analysis, has at least double coverage and consists of 75 overlapping YAC clones, all isolated from CEPH YAC libraries. Their insert sizes were estimated by contour-clamped homogeneous electric field (CHEF) gel electrophoresis. Chromosomal localization and chimerism of the YACs were investigated by FISH analysis. Chimerism of YAC clones was independently determined by restriction mapping. On the basis of YAC end-derived DNA markers and sequence-tagged sites (STSs), with an average spacing of approximately 70 kb, as well as restriction enzyme analysis, a long-range physical map was established for the 6-Mb DNA region of chromosome 12 covered by the YAC contig. Within the YAC contig, the relative positions of various known genes, an expressed sequence-tagged site, and a number of CEPH/Généthon polymorphic markers were determined. The latter data allow full integration of our mapping data with those obtained by CEPH/Généthon as well as those reported at the Second International Workshop on Human Chromosome 12 Mapping. Finally, this YAC contig constitutes the basis for the contstruction of a transcriptional map of this region and is likely to facilitate identification of genes involved in the formation of various benign solid tumor types.
Schoenmakers
E F
EF
Laboratory for Molecular Oncology, Center for Human Genetics, University of Leuven, Belgium.
Geurts
J M
JM
Kools
P F
PF
Mols
R
R
Huysmans
C
C
Bullerdiek
J
J
Van den Berghe
H
H
Van de Ven
W J
WJ
D000236:Adenoma / Q000235:genetics; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002678:Chimera; D002869:Chromosome Aberrations; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast; D002881:Chromosomes, Human, Pair 12; D002885:Chromosomes, Human, Pair 16; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics; D005260:Female; D015723:Gene Library; D005819:Genetic Markers; D006222:Hamartoma / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007889:Leiomyoma / Q000235:genetics; D008067:Lipoma / Q000235:genetics; D008168:Lung / Q000002:abnormalities; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012468:Salivary Gland Neoplasms / Q000235:genetics; D051742:Transcription Factor CHOP; D014157:Transcription Factors / Q000235:genetics; D014178:Translocation, Genetic; D014594:Uterine Neoplasms / Q000235:genetics
D022762:CCAAT-Enhancer-Binding Proteins; C497249:DDIT3 protein, human; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005819:Genetic Markers; D014157:Transcription Factors; D051742:Transcription Factor CHOP