Isolation of two contigs of overlapping cosmids derived from human chromosomal band 3p21.1 and identification of 5 new 3p21.1 genes.
Consistent loss of DNA sequences from several regions on the short arm of human chromosome 3 has suggested that multiple tumor suppressor genes reside on chromosome 3p in various types of cancer cells. We have focused our efforts on an analysis of chromosomal band 3p21.1 since aminoacylase-1 (ACY1), which is localized to this band, has been shown to have lower levels of expression in several small cell and non-small cell lung cancer cell lines. Starting with two cosmids within 3p21.1, D3S92 and D3S93, we have isolated two separate contigs of overlapping cosmids within 3p21.1, by screening a library of 5700 chromosome 3-specific cosmid clones. Detailed restriction maps for these two contigs show that they contain multiple clusters of rare cutting restriction endonuclease sites. One contig extends for 100 kb and encompassed both ACY1 and D3S92, and the other extends about 80 kb around the D3S93 locus. Many different restriction fragments derived from these two contigs were found to be evolutionarily conserved and hybridized to distinct message transcripts. These fragments were used to identify homologous cDNAs from an adenogastric cDNA library, and several of these cDNAs were partially sequenced. We have identified five new genes from these two contigs and there is evidence to suggest that several additional genes reside within these cosmid contigs. The genes identified from 3p21.1 were then hybridized to DNA, isolated from a series of lung cancer cell lines and matched normal and tumor DNA from lung cancer patients. No alterations were detected with any of these probes, both at the DNA or RNA levels. A similar analysis with DNA fragments derived from these two genomic regions also failed to detect any alterations.
Wayne State University, School of Medicine, Detroit, Michigan 48201.
V
V
Shridhar
W
W
Golembieski
A
A
Kamat
S E
SE
Smith
N
N
Phillips
O J
OJ
Miller
Y
Y
Miller
D I
DI
Smith
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002893:Chromosomes, Human, Pair 3 / Q000235:genetics*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003360:Cosmids; D006224:Cricetinae; D018076:DNA, Complementary / Q000302:isolation & purification; D016147:Genes, Tumor Suppressor; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D009693:Nucleic Acid Hybridization; D015183:Restriction Mapping; D017384:Sequence Deletion; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary