Structural basis of sequence-specific Holliday junction cleavage by MOC1

Lin, Huajian; Zhang, Danping; Zuo, Ke; Yuan, Cai; Li, Jinyu; Huang, Mingdong; Lin, Zhonghui

doi:10.1038/s41589-019-0377-4

Cited by 23 publications

(20 citation statements)

References 48 publications

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“…14d). Another study presented the crystal structures of apo and HJ-bound ZmMOC1 50 . The structure of ZmMOC1 in this study is nearly identical to the recently reported ZmMOC1 structure (PDB: 6IS9) with an RMSD of 0.45 Å ( Supplementary Fig.…”

Section: Discussionmentioning

confidence: 99%

Structural insights into sequence-dependent Holliday junction resolution by the chloroplast resolvase MOC1

et al. 2020

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Holliday junctions (HJs) are key DNA intermediates in genetic recombination and are eliminated by nuclease, termed resolvase, to ensure genome stability. HJ resolvases have been identified across all kingdoms of life, members of which exhibit sequence-dependent HJ resolution. However, the molecular basis of sequence selectivity remains largely unknown. Here, we present the chloroplast resolvase MOC1, which cleaves HJ in a cytosine-dependent manner. We determine the crystal structure of MOC1 with and without HJs. MOC1 exhibits an RNase H fold, belonging to the retroviral integrase family. MOC1 functions as a dimer, and the HJ is embedded into the basic cleft of the dimeric enzyme. We characterize a base recognition loop (BR loop) that protrudes into and opens the junction. Residues from the BR loop intercalate into the bases, disrupt the C-G base pairing at the crossover and recognize the cytosine, providing the molecular basis for sequence-dependent HJ resolution by a resolvase.

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Section: Discussionmentioning

confidence: 99%

Structural insights into sequence-dependent Holliday junction resolution by the chloroplast resolvase MOC1

et al. 2020

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“…The selenomethionine (SeMet) labelled protein of Sso2081 was produced as previously described (33). Briefly, the Rosetta (DE3) cells containing pET-Sso2081 plasmid were cultured in Luria-Bertani (LB) media overnight.…”

Section: Methodsmentioning

confidence: 99%

Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081

Luo²,

Lin

2022

Preprint

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The cyclic oligoadenylates (cOAs) act as second messengers of type III CRISPR immunity system through activating the auxiliary nucleases for indiscriminate RNA degradation. The cOA-degrading nucleases (ring nucleases) provide an off-switch regulation of the signaling, thereby preventing cell dormancy or cell death. Here, we describe the crystal structures of the CRISPR-associated ring nuclease 1 (Crn1) from Saccharolobus solfataricus (Sso) 2081 in its apo or bound to cA4 in both pre-cleavage and transient intermediate states. Sso2081 harbors a unique helical insert that encloses cA4 in the central cavity. Two free phosphates symmetrically bind the catalytic site of apo Sso2081 and overlap with the two scissile phosphates of cA4, supporting a bilaterally symmetrical cleavage. The structure of transient intermediate state captured by Ser11Ala mutation immediately illustrates a stepwise cleavage of cA4 by Sso2081. Our study establishes atomic mechanisms of cA4 recognition and degradation by the type III CRISPR ring nuclease Crn1/Sso2081.

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“…XP_033417438.1) was synthesised in Union Biotech (China). Protein expression and purification were performed as previously described 31 . Briefly, The recombinant proteins of Aurora B and INCENP were coexpressed in BL21 (DE3) pLysS competent cells.…”

Section: Methodsmentioning

confidence: 99%

Discovery of a novel Aurora B inhibitor GSK650394 with potent anticancer and anti-aspergillus fumigatus dual efficacies in vitro

et al. 2021

Journal of Enzyme Inhibition and Medicinal Chemistry

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Invasive fungal infections including Candidiasis and Aspergillosis are associated with considerable morbidity and mortality in immunocompromised individuals, such as cancer patients. Aurora B is a key mitotic kinase required for the cell division of eukaryotes from fungus to man. Here, we identified a novel Aurora B inhibitor GSK650394 that can inhibit the recombinant Aurora B from human and Aspergillus fumigatus , with IC 50 values of 5.68 and 1.29 µM, respectively. In HeLa and HepG2 cells, GSK650394 diminishes the endogenous Aurora B activity and causes cell cycle arrest in G2/M phase. Further cell-based assays demonstrate that GSK650394 efficiently suppresses the proliferation of both cancer cells and Aspergillus fumigatus . Finally, the molecular docking calculation and site-directed mutagenesis analyses reveal the molecular mechanism of Aurora B inhibition by GSK650394. Our work is expected to provide new insight into the combinational therapy of cancer and Aspergillus fumigatus infection.

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Structural basis of sequence-specific Holliday junction cleavage by MOC1

Cited by 23 publications

References 48 publications

Structural insights into sequence-dependent Holliday junction resolution by the chloroplast resolvase MOC1

Structural insights into sequence-dependent Holliday junction resolution by the chloroplast resolvase MOC1

Molecular basis of stepwise cyclic tetra-adenylate cleavage by the type III CRISPR ring nuclease Crn1/Sso2081

Discovery of a novel Aurora B inhibitor GSK650394 with potent anticancer and anti-aspergillus fumigatus dual efficacies in vitro

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