Most cancers have multiple chromosomal rearrangements; the molecular mechanisms that generate them remain largely unknown. Mice carrying a heterozygous missense change in one of the DNA-binding domains of Rpa1 develop lymphoid tumors, and their homozygous littermates succumb to early embryonic lethality. Array comparative genomic hybridization of the tumors identified large-scale chromosomal changes as well as segmental gains and losses. The Rpa1 mutation resulted in defects in DNA double-strand break repair and precipitated chromosomal breaks as well as aneuploidy in primary heterozygous mutant mouse embryonic fibroblasts. The equivalent mutation in yeast is hypomorphic and semidominant and enhanced the formation of gross chromosomal rearrangements in multiple genetic backgrounds. These results indicate that Rpa1 functions in DNA metabolism are essential for the maintenance of chromosomal stability and tumor suppression.
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We thank H. Hou, Jr. and B. Jin for technical assistance and S. Brill for providing antibodies. This work was supported by grants from the US National Institutes of Health (to R.K., R.D.K. and W.E.).
The authors declare no competing financial interests.
Molecular dynamics suggests that the L221P mutation disrupts the DNA-binding interface of the oligonucleotide binding (OB) domain 1 of human RPA1. (PDF 5784 kb)
Laser capture microdissection (LCM) and loss of heterozygosity (LOH) analysis. (PDF 379 kb)
Partial dominance of plasmid borne rfa1-L221P alleles for sensitivity to constitutive expression of HO endonuclease. (PDF 7308 kb)
Summary of chromosomal aberrations in lymphomas of Rpa1L230P/+ mice. (PDF 60 kb)
Karyotypic aberrations in Rpa1L230P/+ MEFs. (PDF 51 kb)
Partial dominance of plasmid-borne rfa1-L221P alleles in MMS sensitivity. (PDF 64 kb)
Partial dominance of plasmid-borne rfa1-L221P alleles in HU sensitivity. (PDF 65 kb)
Partial dominance of plasmid-borne rfa1-L221P alleles in UV sensitivity. (PDF 719 kb)
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