TY - JOUR
T1 - siRNA depletion of BRCA1, but not BRCA2, causes increased genome instability in Fanconi anemia cells
AU - Bruun, Donald
AU - Folias, Alexandra
AU - Akkari, Yassmine
AU - Cox, Yumi
AU - Olson, Susan
AU - Moses, Robb
N1 - Funding Information:
We thank A. D’Andrea, M. Grompe, J. Hejna, and S. Moses for providing cell lines, antibody, constructs and discussion. Supported by grant PO1 HL48546 from the US National Institutes of Health.
PY - 2003/9/18
Y1 - 2003/9/18
N2 - BRCA1 and BRCA2 proteins act in repair of interstrand crosslinks (ICLs) and maintenance of genome stability and are known to be part of the Fanconi anemia (FA) pathway. We have investigated the role of the BRCA1 and BRCA2 genes in genome stability following ICL damage in normal and FA cells. To circumvent cell lethality of complete disruptions in BRCA1 or BRCA2, small inhibitory RNA (siRNA) was used to transiently deplete the expression of the proteins. Using chromosomal stability after ICL damage as the end point, we find that BRCA1 functions in more than just the FA pathway for genome maintenance, whereas BRCA2 appears to act predominantly in the FA pathway. Depletion of BRCA1 causes a marked decrease, although not a complete absence of, ubiquitination of FANCD2. In contrast to BRCA1, BRCA2 is not needed for normal ubiquitination of FANCD2 after DNA damage, a requirement for the FA pathway to function. Thus, BRCA2 is epistatic to FA genes for ICL repair, but not for damage-induced modification of FANCD2 and may act downstream form FANCD2.
AB - BRCA1 and BRCA2 proteins act in repair of interstrand crosslinks (ICLs) and maintenance of genome stability and are known to be part of the Fanconi anemia (FA) pathway. We have investigated the role of the BRCA1 and BRCA2 genes in genome stability following ICL damage in normal and FA cells. To circumvent cell lethality of complete disruptions in BRCA1 or BRCA2, small inhibitory RNA (siRNA) was used to transiently deplete the expression of the proteins. Using chromosomal stability after ICL damage as the end point, we find that BRCA1 functions in more than just the FA pathway for genome maintenance, whereas BRCA2 appears to act predominantly in the FA pathway. Depletion of BRCA1 causes a marked decrease, although not a complete absence of, ubiquitination of FANCD2. In contrast to BRCA1, BRCA2 is not needed for normal ubiquitination of FANCD2 after DNA damage, a requirement for the FA pathway to function. Thus, BRCA2 is epistatic to FA genes for ICL repair, but not for damage-induced modification of FANCD2 and may act downstream form FANCD2.
KW - BRCA1
KW - BRCA2
KW - Fanconi anemia
KW - siRNA
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U2 - 10.1016/S1568-7864(03)00112-5
DO - 10.1016/S1568-7864(03)00112-5
M3 - Article
C2 - 12967657
AN - SCOPUS:0041377993
SN - 1568-7864
VL - 2
SP - 1007
EP - 1013
JO - DNA Repair
JF - DNA Repair
IS - 9
ER -