Drosophila damaged DNA binding protein 1 contributes to genome stability in somatic cells.
Drosophila damaged DNA binding protein 1 contributes to genome stability in somatic cells.
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DOI：
10.1093/jb/mvj006
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发表时间：
2006
影响因子：
2.7
通讯作者：
Kaori Shimanouchi;K. Takata;M. Yamaguchi;Shizuka Murakami;Gen Ishikawa;R. Takeuchi;Y. Kanai;Tatsushi Ruike;Ryo-ichi Nakamura;Yoko Abe;K. Sakaguchi
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文献类型：
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作者：
Kaori Shimanouchi;K. Takata;M. Yamaguchi;Shizuka Murakami;Gen Ishikawa;R. Takeuchi;Y. Kanai;Tatsushi Ruike;Ryo-ichi Nakamura;Yoko Abe;K. Sakaguchi
受损的DNA结合蛋白(DDB)复合体由p127(DDB1)和P48(DDB2)亚基组成，被认为在核苷酸切除修复(NER)中起作用。我们利用GAL4-UAS靶向表达系统敲除果蝇翅膀想象盘中的DDB1。通过过量表达D-DDB1基因的反向重复RNA[UAS-D-DDB1(650)-dsRNA]，在转基因果蝇中实现了基因敲除。由于RNA干扰(RNAi)，苍蝇的翅膀表型缩小。翼斑点试验表明，在翼成像盘中D-DDB1的RNAi敲除后，转基因果蝇的基因组不稳定。用化学诱变剂甲基甲烷磺酸(MMS)处理D-DDB1基因RNAi敲除的果蝇幼虫时，与未处理的转基因果蝇相比，具有严重缩小翅膀表型的果蝇的频率增加。这些结果表明，DDB1在果蝇体细胞对MMS损伤的DNA的应答和基因组的稳定性中起着重要作用。
The damaged DNA-binding protein (DDB) complex consists of a heterodimer of p127 (DDB1) and p48 (DDB2) subunits and is believed to have a role in nucleotide excision repair (NER). We used the GAL4-UAS targeted expression system to knock down DDB1 in wing imaginal discs of Drosophila. The knock-down was achieved in transgenic flies using over-expression of inverted repeat RNA of the D-DDB1 gene [UAS-D-DDB1(650)-dsRNA]. As a consequence of RNA interference (RNAi), the fly had a shrunken wing phenotype. The wing spot test showed induced genome instability in transgenic flies with RNAi knock-down of D-DDB1 in wing imaginal discs. When Drosophila larvae with RNAi knock-down of D-DDB1 in wing imaginal discs were treated with the chemical mutagen methyl methanesulfonate (MMS), the frequency of flies with a severely shrunken wing phenotype increased compared to non-treated transgenic flies. These results suggested that DDB1 plays a role in the response to DNA damaged with MMS and in genome stability in Drosophila somatic cells.