Structural insights into recognition and repair of UV-DNA damage by Spore Photoproduct Lyase, a radical SAM enzyme

被引:65
作者
Benjdia, Alhosna [1 ]
Heil, Korbinian [2 ]
Barends, Thomas R. M. [1 ]
Carell, Thomas [2 ]
Schlichting, Ilme [1 ]
机构
[1] Max Planck Inst Med Res, Dept Biomol Mech, D-69120 Heidelberg, Germany
[2] Univ Munich, Dept Chem, Ctr Integrated Prot Sci CiPSM, D-81377 Munich, Germany
关键词
CRYSTAL-STRUCTURE; BACILLUS-SUBTILIS; MECHANISM; LESION; PHOTOLYASE; LIGHT; 5-THYMINYL-5,6-DIHYDROTHYMINE; SUPERFAMILY; REDUCTASE; ALIGNMENT;
D O I
10.1093/nar/gks603
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Bacterial spores possess an enormous resistance to ultraviolet (UV) radiation. This is largely due to a unique DNA repair enzyme, Spore Photoproduct Lyase (SP lyase) that repairs a specific UV-induced DNA lesion, the spore photoproduct (SP), through an unprecedented radical-based mechanism. Unlike DNA photolyases, SP lyase belongs to the emerging superfamily of radical S-adenosyl-l-methionine (SAM) enzymes and uses a [4Fe-4S](1+) cluster and SAM to initiate the repair reaction. We report here the first crystal structure of this enigmatic enzyme in complex with its [4Fe-4S] cluster and its SAM cofactor, in the absence and presence of a DNA lesion, the dinucleoside SP. The high resolution structures provide fundamental insights into the active site, the DNA lesion recognition and binding which involve a beta-hairpin structure. We show that SAM and a conserved cysteine residue are perfectly positioned in the active site for hydrogen atom abstraction from the dihydrothymine residue of the lesion and donation to the alpha-thyminyl radical moiety, respectively. Based on structural and biochemical characterizations of mutant proteins, we substantiate the role of this cysteine in the enzymatic mechanism. Our structure reveals how SP lyase combines specific features of radical SAM and DNA repair enzymes to enable a complex radical-based repair reaction to take place.
引用
收藏
页码:9308 / 9318
页数:11
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