Functional organization of box C/D RNA-guided RNA methyltransferase

TY - JOUR

T1 - Functional organization of box C/D RNA-guided RNA methyltransferase

AU - Yang, Zuxiao

AU - Wang, Jiayin

AU - Huang, Lin

AU - Lilley, David M. J.

AU - Ye, Keqiong

N1 - National Natural Science Foundation of China [91940302, 91540201, 31430024, 31325007]; National Key R&D Program of China [2017YFA0504600]; Strategic Priority Research Program of Chinese Academy of Sciences [XDB08010203]; 100 Talents Program of CAS. Funding for open access charge: National Natural Science Foundation of China. © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2020/5/21

Y1 - 2020/5/21

N2 - Box C/D RNA protein complexes (RNPs) catalyze site-specific 2'-O-methylation of RNA with specificity determined by guide RNAs. In eukaryotic C/D RNP, the paralogous Nop58 and Nop56 proteins specifically associate with terminal C/D and internal C'/D' motifs of guide RNAs, respectively. We have reconstituted active C/D RNPs with recombinant proteins of the thermophilic yeast Chaetomium thermophilum. Nop58 and Nop56 could not distinguish between the two C/D motifs in the reconstituted enzyme, suggesting that the assembly specificity is imposed by trans-acting factors in vivo. The two C/D motifs are functionally independent and halfmer C/D RNAs can also guide site-specific methylation. Extensive pairing between C/D RNA and substrate is inhibitory to modification for both yeast and archaeal C/D RNPs. N6-methylated adenine at box D/D' interferes with the function of the coupled guide. Our data show that all C/D RNPs share the same functional organization and mechanism of action and provide insight into the assembly specificity of eukaryotic C/D RNPs.

AB - Box C/D RNA protein complexes (RNPs) catalyze site-specific 2'-O-methylation of RNA with specificity determined by guide RNAs. In eukaryotic C/D RNP, the paralogous Nop58 and Nop56 proteins specifically associate with terminal C/D and internal C'/D' motifs of guide RNAs, respectively. We have reconstituted active C/D RNPs with recombinant proteins of the thermophilic yeast Chaetomium thermophilum. Nop58 and Nop56 could not distinguish between the two C/D motifs in the reconstituted enzyme, suggesting that the assembly specificity is imposed by trans-acting factors in vivo. The two C/D motifs are functionally independent and halfmer C/D RNAs can also guide site-specific methylation. Extensive pairing between C/D RNA and substrate is inhibitory to modification for both yeast and archaeal C/D RNPs. N6-methylated adenine at box D/D' interferes with the function of the coupled guide. Our data show that all C/D RNPs share the same functional organization and mechanism of action and provide insight into the assembly specificity of eukaryotic C/D RNPs.

UR - https://www.scopus.com/pages/publications/85084943415

U2 - 10.1093/nar/gkaa247

DO - 10.1093/nar/gkaa247

M3 - Article

C2 - 32297938

SN - 0305-1048

VL - 48

SP - 5094

EP - 5105

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 9

ER -