A Type III-B Cmr effector complex catalyzes the synthesis of cyclic oligoadenylate second messengers by cooperative substrate binding

Nucleic Acids Research
Wenyuan HanQunxin She

Abstract

Recently, Type III-A CRISPR-Cas systems were found to catalyze the synthesis of cyclic oligoadenylates (cOAs), a second messenger that specifically activates Csm6, a Cas accessory RNase and confers antiviral defense in bacteria. To test if III-B CRISPR-Cas systems could mediate a similar CRISPR signaling pathway, the Sulfolobus islandicus Cmr-α ribonucleoprotein complex (Cmr-α-RNP) was purified from the native host and tested for cOA synthesis. We found that the system showed a robust production of cyclic tetra-adenylate (c-A4), and that c-A4 functions as a second messenger to activate the III-B-associated RNase Csx1 by binding to its CRISPR-associated Rossmann Fold domain. Investigation of the kinetics of cOA synthesis revealed that Cmr-α-RNP displayed positively cooperative binding to the adenosine triphosphate (ATP) substrate. Furthermore, mutagenesis of conserved domains in Cmr2α confirmed that, while Palm 2 hosts the active site of cOA synthesis, Palm 1 domain serves as the primary site in the enzyme-substrate interaction. Together, our data suggest that the two Palm domains cooperatively interact with ATP molecules to achieve a robust cOA synthesis by the III-B CRISPR-Cas system.

References

Jul 15, 1987·European Journal of Biochemistry·J Ricard, A Cornish-Bowden
Oct 1, 1980·Archives of Biochemistry and Biophysics·A E Oleson, M Sasakuma
Dec 5, 2002·Cell·Marc Dreyfus, Philippe Régnier
Dec 20, 2008·Science·Luciano A Marraffini, Erik J Sontheimer
Dec 1, 2009·Cell·Caryn R HaleMichael P Terns
Jan 15, 2010·Nature·Luciano A Marraffini, Erik J Sontheimer
Mar 13, 2012·Structure·Alexis I CocozakiHong Li
Jan 17, 2013·Molecular Microbiology·Ling DengQunxin She
Apr 16, 2013·Journal of Molecular Biology·Takuo OsawaTomoyuki Numata
Jul 12, 2013·PLoS Computational Biology·Melanie I Stefan, Nicolas Le Novère
Nov 5, 2013·Journal of Bacteriology·Asma Hatoum-AslanLuciano A Marraffini
Feb 18, 2014·RNA Biology·Gisle VestergaardShiraz A Shah
Apr 15, 2014·Nucleic Acids Research·Krzysztof ChylinskiEugene V Koonin
May 13, 2014·Frontiers in Genetics·Kira S MakarovaL Aravind
Oct 19, 2014·Nucleic Acids Research·Linlin HouElena Evguenieva-Hackenberg
Nov 5, 2014·Genes & Development·Caryn R HaleMichael P Terns
Dec 3, 2014·Molecular Cell·Raymond H J StaalsJohn van der Oost
Dec 3, 2014·Molecular Cell·Gintautas TamulaitisVirginijus Siksnys
Dec 3, 2014·Journal of Molecular Biology·Maycon C OliveiraChuck S Farah
Dec 30, 2014·Nucleic Acids Research·Xing Zhu, Keqiong Ye
May 3, 2015·FEMS Microbiology Reviews·André PlagensLennart Randau
May 12, 2015·Cell·Poulami SamaiLuciano A Marraffini
May 20, 2015·Methods in Molecular Biology·Jing Zhang, Malcolm F White
May 23, 2015·FEMS Microbiology Reviews·Emmanuelle CharpentierMalcolm F White
Sep 29, 2015·Nature Reviews. Microbiology·Kira S MakarovaEugene V Koonin
Oct 4, 2015·Nature·Luciano A Marraffini
Jan 12, 2016·Nature Reviews. Microbiology·Gil Amitai, Rotem Sorek
Jan 24, 2016·Nucleic Acids Research·Jing ZhangMalcolm F White
Feb 6, 2016·Genes & Development·Joshua R ElmoreMichael P Terns
Feb 6, 2016·Genes & Development·Michael A EstrellaScott Bailey
Mar 8, 2016·Molecular Cell·Samuel H SternbergUdi Qimron
Aug 6, 2016·Science·Prarthana MohanrajuJohn van der Oost
Oct 25, 2016·Trends in Microbiology·Gintautas TamulaitisVirginijus Siksnys
Apr 8, 2017·Science·Simon A JacksonStan J J Brouns

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Citations

May 22, 2019·The Journal of Biological Chemistry·Kaitlin JohnsonScott Bailey
Nov 12, 2019·Frontiers in Microbiology·Yingjun Li, Nan Peng
Aug 21, 2020·The Journal of Biological Chemistry·Tina Y Liu, Jennifer A Doudna
Dec 14, 2018·Emerging Topics in Life Sciences·Yan ZhangQunxin She
Feb 9, 2021·Frontiers in Microbiology·Fengtao Huang, Bin Zhu

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Methods Mentioned

BETA
PCR
size exclusion chromatography
electrophoresis
cleavage assay
electrophoretic mobility shift assay

Software Mentioned

OriginLab
OriginPro
ImageQuant

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