Molecular cloning of the papillary renal cell carcinoma-associated translocation (X;1)(p11;q21) breakpoint.
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.
Weterman
M A
MA
Department of Human Genetics, University Hospital Nijmegen, The Netherlands.
Wilbrink
M
M
Janssen
I
I
Janssen
H A
HA
van den Berg
E
E
Fisher
S E
SE
Craig
I
I
Geurts van Kessel
A
A
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002291:Carcinoma, Papillary / Q000235:genetics*; D002292:Carcinoma, Renal Cell / Q000235:genetics*; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast; D002878:Chromosomes, Human, Pair 1; D003001:Cloning, Molecular; D003360:Cosmids; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D005819:Genetic Markers; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007680:Kidney Neoplasms / Q000235:genetics*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D014178:Translocation, Genetic; D014960:X Chromosome
D018076:DNA, Complementary; D004273:DNA, Neoplasm; D005819:Genetic Markers