کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1980279 | 1061838 | 2013 | 11 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: A proteome-wide visual screen identifies fission yeast proteins localizing to DNA double-strand breaks A proteome-wide visual screen identifies fission yeast proteins localizing to DNA double-strand breaks](/preview/png/1980279.png)
• We used a fission yeast ORFeome library to screen for proteins localizing to DSBs.
• We identified 51 proteins that can form a nuclear focus at an HO-induced DSB.
• We characterized the focus emergence timing of DSB-localizing proteins.
• We analyzed the roles of two novel DSB-localizing proteins identified in our screen.
DNA double-strand breaks (DSBs) are a major threat to genome integrity. Proteins involved in DNA damage checkpoint signaling and DSB repair often relocalize and concentrate at DSBs. Here, we used an ORFeome library of the fission yeast Schizosaccharomyces pombe to systematically identify proteins targeted to DSBs. We found 51 proteins that, when expressed from a strong exogenous promoter on the ORFeome plasmids, were able to form a distinct nuclear focus at an HO endonuclease-induced DSB. The majority of these proteins have known connections to DNA damage response, but few have been visualized at a specific DSB before. Among the screen hits, 37 can be detected at DSBs when expressed from native promoters. We classified them according to the focus emergence timing of the endogenously tagged proteins. Eight of these 37 proteins are yet unnamed. We named these eight proteins DNA-break-localizing proteins (Dbls) and performed preliminary functional analysis on two of them, Dbl1 (SPCC2H8.05c) and Dbl2 (SPCC553.01c). We found that Dbl1 and Dbl2 contribute to the normal DSB targeting of checkpoint protein Rad26 (homolog of human ATRIP) and DNA repair helicase Fml1 (homolog of human FANCM), respectively. As the first proteome-wide inventory of DSB-localizing proteins, our screen result will be a useful resource for understanding the mechanisms of eukaryotic DSB response.
Journal: DNA Repair - Volume 12, Issue 6, 1 June 2013, Pages 433–443