Construction of a consistent YAC contig for human chromosome region 3p14.1.

Abstract

Chromosomal deletions and translocations of human chromosome region 3pl4 are observed in various human malignancies and suggest the existence of a tumor suppressor gene locus within this region. Tumors most frequently affected by these aberrations are small-cell lung cancer and renal-cell carcinoma. In continuation of our previously published YAC contig of chromosome region 3p14.2-p14.3, we report here on the construction of a YAC contig of at least II Mb that consisted of 171 YACs and covers the entire subregion 3pl4.1. This contig includes the t(3;8) breakpoint of a hereditary renal-cell carcinoma localized in 3pl4.2 and extends into human chromosome region 3p12-p13. it defines the order of 34 DNA probes in relation to reference markers D3S6 and D3S30 as well as the human protein tyrosine phosphatase-~ gene. For 31 DNA probes we identified nonchimeric YACs by fluorescence in situ hybridization. The minimal tiling pathway consists of 16 yeast artificial chromosomes. As a prerequisite for identification of a putative tumor suppressor gene within this region, this contig renders human chromosome region 3pl4.1 accessible to gene isolation.

The p44S10 locus, encoding a subunit of the proteasome regulatory particle, is amplified during progression of cutaneous malignant melanoma

“…A SCH monochromosome mapping panel localized the gene to chromosome 3; a SCH panel containing di erent portions of chromosome 3 localized the gene to 3p14.1-2; contiguous YAC clones spanning 3p14 mapped the gene to the speci®c CEPH YACs (Figure 2). Positive clones included 371h4, 879h10, 927e1, 924b12, 882d9, 965h3, 936c10, 977c8, 846g11, 403b2, and 965a3, and no discordances with mapped YACs were observed (Bardenheuer et al, 1996). The p44 S10 locus in 3p14.1 is approximately 1 ± 2.7 Mb proximal to the t3;8 translocation breakpoint, as de®ned by alignment of non-overlapping and nonchimeric YACs (144g3,166g8,130h11,850a6).…”

Minimal deletion of 3p13→14.2 associated with immortalization of human uroepithelial cells

“…The dotted line (//---//) region is not drawn to scale. Selected markers are U2020d, distal breakpoint of the U2020 cell line deletion; PTPRG, protein tyrosine phosphatase gamma locus; the FRA3B fragile site; the t(3;8) translocation breakpoint; and FHIT, fragile histidine triad locus Michaelis et al, 1995;Todd et al, 1995;Bardenheuer et al, 1996). was highly significant (P ϭ 7 ϫ 10 Ϫ6 ).…”

“…3A), FISH analyses were performed using YAC probes (Fig. 3B) which map specifically to 3p13=14.3 Michaelis et al, 1995;Todd et al, 1995;Bardenheuer et al, 1996). The centromeric breakpoint in the smallest common region deleted was defined cytogenetically as 3p14.1 using ␦E6-1 (Figs.…”

Identification of novel 'expressed sequence tags' within the FHIT gene locus in human chromosome region 3p14.2

“…1) was chosen for direct cDNA selection. This YAC has been shown to be non-chimeric by fluorescence in situ hybridization (Michaelis et al 1995) and is located distal to the t(3;8) translocation breakpoint and the FRA3B fragile site (Gemmill et al 1995;Aburatani et al 1996;Bardenheuer et al 1996). It is flanked by markers D3S1300/BE758-6 and D3S1313 which show a high frequency of allelic loss in renal cell carcinoma (Lubinski et al 1994;Wilhelm et al 1995;Willers et al 1996).…”

“…Chromosomal assignment of the 11 unique clones was performed by PCR analyses and Southern blot hybridizations using DNA from human placenta, CHO cells, cell line 314-1b, yeast strain AB1380, YAC 268F1, as well as several other YACs from our 3p14-specific YAC contig (Michaelis et al 1995;Bardenheuer et al 1996) Table 1. The localization of these clones within our previously published 3p14-specific YAC contig is shown in Fig.…”