SiRNA depletion of BRCA1, but not BRCA2, causes increased genome instability in Fanconi anemia cells
SiRNA depletion of BRCA1, but not BRCA2, causes increased genome instability in Fanconi anemia cells
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DOI：
10.1016/s1568-7864(03)00112-5
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发表时间：
2003-09-18
期刊：
影响因子：
3.8
通讯作者：
Moses, R
中科院分区：
文献类型：
--
作者：
Bruun, D;Folias, A;Moses, R
BRCA1和BRCA2蛋白在修复链间交联（ICL）和维持基因组稳定性中起作用，已知是范可尼贫血（FA）途径的一部分。我们研究了BRCA1和BRCA2基因在正常和FA细胞ICL损伤后基因组稳定性中的作用。为了避免BRCA 1或BRCA 2完全破坏的细胞致死性，使用小抑制性RNA（siRNA）瞬时耗尽蛋白质的表达。使用ICL损伤后的染色体稳定性作为终点，我们发现BRCA1的功能不仅仅是在FA通路中维持基因组，而BRCA2似乎主要在FA通路中起作用。BRCA 1的消耗导致FANCD 2的泛素化显著减少，尽管不是完全缺失。与BRCA 1相反，BRCA 2在DNA损伤后不需要FANCD 2的正常泛素化，这是FA途径发挥作用的必要条件。因此，BRCA 2是上位性的ICL修复FA基因，但不是损伤诱导的FANCD 2的修饰，并可能从FANCD 2的下游。(C)2003 Elsevier B.V保留所有权利。
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. (C) 2003 Elsevier B.V All rights reserved.