Molecular cloning of the papillary renal cell carcinoma-associated translocation (X;1)(p11;q21) breakpoint

• Published in: Cytogenetics and cell genetics Vol. 75; no. 1; p. 2
• Main Authors: Weterman, M A, Wilbrink, M, Janssen, I, Janssen, H A, van den Berg, E, Fisher, S E, Craig, I, Geurts van Kessel, A
• Format: Journal Article
• Language: English
• Published: Switzerland 1996
• Subjects: Animals; Base Sequence; Blotting, Southern; Carcinoma, Papillary - genetics; Carcinoma, Renal Cell - genetics; Chromosome Mapping; Chromosomes, Artificial, Yeast; Chromosomes, Human, Pair 1; Cloning, Molecular; Cosmids; DNA, Complementary - genetics; DNA, Neoplasm - genetics; Genetic Markers; Humans; In Situ Hybridization, Fluorescence; Kidney Neoplasms - genetics; Male; Mice; Molecular Sequence Data; Translocation, Genetic; X Chromosome
• ISSN: 0301-0171
• DOI: 10.1159/000134444

A combination of Southern blot analysis on a panel of tumor-derived somatic cell hybrids and fluorescence in situ hybridization techniques was used to map YACs, cosmids and DNA markers from the Xp11.2 region relative to the X chromosome breakpoint of the renal cell carcinoma-associated t(X;1)(p11;q21). The position of the breakpoint could be determined as follows: Xcen-OATL2-DXS146-DXS255-SYP-t(X;1)-TFE 3-OATL1-Xpter. Fluorescence in situ hybridization experiments using TFE3-containing YACs and cosmids revealed split signals indicating that the corresponding DNA inserts span the breakpoint region. Subsequent Southern blot analysis showed that a 2.3-kb EcoRI fragment which is present in all TFE3 cosmids identified, hybridizes to aberrant restriction fragments in three independent t(X;1)-positive renal cell carcinoma DNAs. The breakpoints in these tumors are not the same, but map within a region of approximately 6.5 kb. Through preparative gel electrophoresis an (X;1) chimaeric 4.4-kb EcoRI fragment could be isolated which encompasses the breakpoint region present on der(X). Preliminary characterization of this fragment revealed the presence of a 150-bp region with a strong homology to the 5' end of the mouse TFE3 cDNA in the X-chromosome part, and a 48-bp segment in the chromosome 1-derived part identical to the 5' end of a known EST (accession number R93849). These observations suggest that a fusion gene is formed between the two corresponding genes in t(X;1)(p11;q21)-positive papillary renal cell carcinomas.