A Type I-F Anti-CRISPR Protein Inhibits the CRISPR-Cas Surveillance Complex by ADP-Ribosylation

Niu, Yiying; Yang, Lingguang; Gao, Teng; Dong, Changpeng; Zhang, Buyu; Yin, Peng; Hopp, Ann-Katrin; Li, Dongdong; Gan, Rui; Wang, Hongou; Liu, Xi; Cao, Xueli; Xie, Ying; Meng, Xianbin; Deng, Haiteng; Zhang, Xiaohui; Ren, Jie; Hottiger, Michael O.; Chen, Zeliang; Zhang, Yi; Liu, Xiaoyun; Feng, Yue

doi:10.1016/j.molcel.2020.09.015

Cited by 44 publications

(25 citation statements)

References 71 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the Csy–AcrIF14–dsDNA SP structure, consistent with previous studies ( 5 , 7 ), K247 and N250 of the Cas8f subunit are involved in dsDNA SP binding (Figure 5B ). In our previous study, we also confirmed that the K247E and N250D mutations of the Cas8f subunit both abolished the dsDNA SP binding of the Csy complex ( 14 ). To investigate the roles of these two residues in dsDNA binding by the Csy–AcrIF14 complex, we purified Csy–AcrIF14 complexes in which the Csy complex included either the Cas8f K247E or the Cas8f N250D mutation.…”

Section: Resultssupporting

confidence: 83%

“…In our previous study ( 14 ), we have proved that AcrIF14 interacts with the Csy complex, suggesting that it may achieve its inhibition through the interaction. To determine the interaction mode between AcrIF14 and the Csy complex, we prepared the Csy–AcrIF14 complex and determined the structure using cryo-electron microscopy (cryo-EM) at a resolution of 3.43 Å (Figures 2 and Supplementary Figure S2 , and Supplementary Table S2 ).…”

Section: Resultsmentioning

confidence: 75%

“…The full-length AcrIF9 gene was also synthesized by GenScript and cloned into pET28a to produce a His-tagged fusion protein. The GST-tagged and His-tagged proteins were purified as described ( 14 ), respectively. Briefly, the proteins were expressed in Escherichia coli strain BL21 and induced by 0.2 mM isopropyl-β- d -thiogalactopyranoside (IPTG) when the cell density reached an OD 600 nm of 0.8.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, another 10 type I-F Acr genes were uncovered, ranging from AcrIF15 to AcrIF24, rendering the type I-F Acr family the largest Acr family ( 17 ). The structural basis of the mechanism of type I-F Acrs has been determined for AcrIF1 and AcrIF2 ( 4–6 ), AcrIF3 ( 7 , 18 , 19 ), AcrIF6/8/9 ( 20 , 21 ), AcrIF7 ( 22 ), AcrIF10 ( 5 ) and AcrIF11 ( 14 ). A canonical suppression strategy of Acr is sterically blocking the hybridization between the complementary DNA strand and the crRNA.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Insights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR–Cas surveillance complex

Liu

Zhang

Xiu

et al. 2021

Nucleic Acids Research

Self Cite

View full text Add to dashboard Cite

CRISPR–Cas systems are bacterial adaptive immune systems, and phages counteract these systems using many approaches such as producing anti-CRISPR (Acr) proteins. Here, we report the structures of both AcrIF14 and its complex with the crRNA-guided surveillance (Csy) complex. Our study demonstrates that apart from interacting with the Csy complex to block the hybridization of target DNA to the crRNA, AcrIF14 also endows the Csy complex with the ability to interact with non-sequence-specific dsDNA as AcrIF9 does. Further structural studies of the Csy–AcrIF14–dsDNA complex and biochemical studies uncover that the PAM recognition loop of the Cas8f subunit of the Csy complex and electropositive patches within the N-terminal domain of AcrIF14 are essential for the non-sequence-specific dsDNA binding to the Csy–AcrIF14 complex, which is different from the mechanism of AcrIF9. Our findings highlight the prevalence of Acr-induced non-specific DNA binding and shed light on future studies into the mechanisms of such Acr proteins.

show abstract

Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 75%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Insights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR–Cas surveillance complex

Liu

Zhang

Xiu

et al. 2021

Nucleic Acids Research

Self Cite

View full text Add to dashboard Cite

show abstract

“…CRISPR-Cas systems are divided into two distinct classes, class I and II, which are classified by multi-protein or single-protein Cas effector and further divided into six types (type I-VI) (6). The type I-F CRISPR-Cas surveillance complex, also called the Csy complex, consists of four types of Cas subunits including Cas8f, Cas5f, Cas7f and Cas6f (Figure S1A), in a stoichiometry of 1:1:6:1, and is guided by a ~60-nt crRNA (Figure S1B) (3,7,8). After the Csy complex binds the target DNA and forms an R-loop structure of the DNA, the Cas2/3 nuclease is recruited to the Csy-DNA complex and further degrades the DNA molecule (9,10).…”

Section: Introductionmentioning

confidence: 99%

Mechanistic insights into the inhibition of the CRISPR-Cas surveillance complex by anti-CRISPR protein AcrIF13

Wang

Gao

Zhou

et al. 2022

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

show abstract

Legionella pneumophila temporally regulates the activity of ADP/ATP translocases by reversible ADP‐ribosylation

Guan

et al. 2022

mLife

View full text Add to dashboard Cite

The mitochondrion is an important signaling hub that governs diverse cellular functions, including metabolism, energy production, and immunity. Among the hundreds of effectors translocated into host cells by the Dot/Icm system of Legionella pneumophila, several are targeted to mitochondria but the function of most of them remains elusive. Our recent study found that the effector Ceg3 inhibits the activity of ADP/ATP translocases (ANTs) by ADP‐ribosylation (ADPR). Here, we show that the effect of Ceg3 is antagonized by Larg1, an effector encoded by lpg0081, a gene that is situated next to ceg3. Larg1 functions to reverse Ceg3‐mediated ADPR of ANTs by cleaving the N‐glycosidic bond between the ADPR moiety and the modified arginine residues in ANTs, leading to restoration of their activity in ADP/ATP exchange. Structural analysis of Larg1 and its complex with ADPR reveals that this ADPR glycohydrolase harbors a unique macrodomain that catalyzes the removal of ADPR modification on ANTs. Our results also demonstrate that together with Ceg3, Larg1 imposes temporal regulation of the activity of ANTs by reversible ADPR during L. pneumophila infection.

show abstract

A Type I-F Anti-CRISPR Protein Inhibits the CRISPR-Cas Surveillance Complex by ADP-Ribosylation

Cited by 44 publications

References 71 publications

Insights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR–Cas surveillance complex

Insights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR–Cas surveillance complex

Mechanistic insights into the inhibition of the CRISPR-Cas surveillance complex by anti-CRISPR protein AcrIF13

Legionella pneumophila temporally regulates the activity of ADP/ATP translocases by reversible ADP‐ribosylation

Contact Info

Product

Resources

About