A 2-Mb YAC contig and physical map covering the chromosome 8q12 breakpoint cluster region in pleomorphic adenomas of the salivary glands.
Pleomorphic adenomas are benign epithelial tumors originating from the major and minor salivary glands. Extensive cytogenetic studies have demonstrated that they frequently show chromosome abnormalities involving chromosome 8, with consistent breakpoints at 8q12. In previous studies, we have shown that these breakpoints are located in a 9-cM interval between MOS/D8S285 and D8S260. Here, we describe directional chromosome walking studies starting from D8S260 as well as D8S285. Using the CEPH and ICRF YAC libraries, these studies resulted in the construction of two nonoverlapping YAC contigs of about 2 and 5 Mb, respectively. Initial fluorescence in situ hybridization (FISH) analysis suggested that the majority of 8q12 breakpoints clustered within the 2-Mb contig, which was mapped to the centromeric part of chromosome band 8q12. This contig has at least double coverage and consists of 34 overlapping YAC clones. The localization of the YACs was confirmed by FISH analysis. On the basis of mapping data of landmarks with an average spacing of 65 kb as well as restriction enzyme analysis, a long-range physical map was established for the chromosome region spanned by the 2-Mb contig. The relative positions of various known genes and expressed sequence tags within this contig were also determined. Subsequent FISH analyses of pleomorphic adenomas using YACs as well as cosmids revealed that all but two of the 8q12 breakpoints in the primary tumors tested mapped within a 300-kb interval between the MOS proto-oncogene and STS EM156. The target gene affected by the chromosome aberrations mapping within this interval was recently shown to be the PLAG1 gene, which encodes a novel zinc finger protein.
Kas
K
K
Center for Human Genetics, University of Leuven, and Flanders Interuniversity Institute for Biotechnology, Belgium.
Röijer
E
E
Voz
M
M
Meyen
E
E
Stenman
G
G
Van de Ven
W J
WJ
D008949:Adenoma, Pleomorphic / Q000235:genetics*; D002503:Centromere / Q000235:genetics; D002873:Chromosome Fragility; D002874:Chromosome Mapping; D016386:Chromosome Walking / Q000379:methods; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D002898:Chromosomes, Human, Pair 8 / Q000235:genetics*; D018899:CpG Islands / Q000235:genetics; D005796:Genes / Q000235:genetics / Q000502:physiology; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D011110:Polymorphism, Genetic; D000090063:Proto-Oncogene Mas; D015183:Restriction Mapping / Q000379:methods; D012468:Salivary Gland Neoplasms / Q000235:genetics*; D012469:Salivary Glands / Q000737:chemistry / Q000473:pathology; D016324:Sequence Tagged Sites
D005819:Genetic Markers; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas