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Researchers Reveal Molecular Mechanism of Type III CRISPR-Cas System
Dec 01, 2018
Type III CRISPR-Cas systems are characterized by the presence of Cas10 protein and are subdivided into four (III A-D) subtypes.
Types III-A system comprises a Csm effector complex which contains five different Csm proteins (Csm1-5) and one CRISPR RNA (crRNA). The crRNA-guided complexes recognize complementary RNA targets and cleave them via the Csm3 subunit.
In a recent study published in Cell, Prof. WANG Yanli's group and Prof. ZHANG XinZheng's group at Institute of Biophysics of Chinese Academy of Sciences reported the crystal structure of Csm and a series of cryo-EM structures of Csm in complex either with cognate or non-cognate RNA targets.
They revealed that Csm complex is composed of five Csm subunits (Csm1-5) and one crRNA with a protein stoichiometry of Csm1122334151.
The spacer region of the crRNA form base was revealed to pair with the complementary target RNA, forming crRNA-target RNA duplex with every sixth base being flipped out, and Csm3 proteins was showed to cleave the target RNA periodically.
By comparing cognate RNA and non-cognate RNA bound Csm complexes, researchers showed that the 3' anti-tag region of target RNA has two distinct binding channels depending on complementarity between the 5'-tag of crRNA and 3' anti-tag of target RNA.
Csm1 subunit undergoes a conformational change upon the cognate target RNA binding, allosterically activating the DNA cleavage and cOA synthesis.
The results provided insights into the mechanistic processes required for both crRNA-meditated, sequence-specific RNA cleavage, as well as transcription-dependent, non-specific DNA cleavage and cOAs generation, which will facilitate the research on type III CRISPR-Cas system.
CRISPR-Cas are RNA-guided adaptive immune systems that protect most archaea and approximately half of bacteria against invading foreign nucleic acids.
All the cryo electron microscopy work was performed in Center for Biological Imaging, Institute of Biophysics of Chinese Academy of Sciences. The X-ray diffraction data were collected at the BL-17U1, and BL-19U1 beamlines at the Shanghai Synchrotron Radiation Facility.
Researchers Reveal Molecular Mechanism of Type III CRISPR-Cas System---Chinese Academy of Sciences
Dec 01, 2018
Type III CRISPR-Cas systems are characterized by the presence of Cas10 protein and are subdivided into four (III A-D) subtypes.
Types III-A system comprises a Csm effector complex which contains five different Csm proteins (Csm1-5) and one CRISPR RNA (crRNA). The crRNA-guided complexes recognize complementary RNA targets and cleave them via the Csm3 subunit.
In a recent study published in Cell, Prof. WANG Yanli's group and Prof. ZHANG XinZheng's group at Institute of Biophysics of Chinese Academy of Sciences reported the crystal structure of Csm and a series of cryo-EM structures of Csm in complex either with cognate or non-cognate RNA targets.
They revealed that Csm complex is composed of five Csm subunits (Csm1-5) and one crRNA with a protein stoichiometry of Csm1122334151.
The spacer region of the crRNA form base was revealed to pair with the complementary target RNA, forming crRNA-target RNA duplex with every sixth base being flipped out, and Csm3 proteins was showed to cleave the target RNA periodically.
By comparing cognate RNA and non-cognate RNA bound Csm complexes, researchers showed that the 3' anti-tag region of target RNA has two distinct binding channels depending on complementarity between the 5'-tag of crRNA and 3' anti-tag of target RNA.
Csm1 subunit undergoes a conformational change upon the cognate target RNA binding, allosterically activating the DNA cleavage and cOA synthesis.
The results provided insights into the mechanistic processes required for both crRNA-meditated, sequence-specific RNA cleavage, as well as transcription-dependent, non-specific DNA cleavage and cOAs generation, which will facilitate the research on type III CRISPR-Cas system.
CRISPR-Cas are RNA-guided adaptive immune systems that protect most archaea and approximately half of bacteria against invading foreign nucleic acids.
All the cryo electron microscopy work was performed in Center for Biological Imaging, Institute of Biophysics of Chinese Academy of Sciences. The X-ray diffraction data were collected at the BL-17U1, and BL-19U1 beamlines at the Shanghai Synchrotron Radiation Facility.
Researchers Reveal Molecular Mechanism of Type III CRISPR-Cas System---Chinese Academy of Sciences