TY - JOUR
T1 - The mechanism of the nucleo-sugar selection by multi-subunit RNA polymerases
AU - Mäkinen, Janne J.
AU - Shin, Yeonoh
AU - Vieras, Eeva
AU - Virta, Pasi
AU - Metsä-Ketelä, Mikko
AU - Murakami, Katsuhiko S.
AU - Belogurov, Georgiy A.
N1 - Funding Information:
We thank Irina Artsimovitch for critically reading the manuscript, the staff at the MacCHESS for support of crystallographic data collection, Anssi M. Malinen for constructing plasmids, and Matti Turtola for his contribution to the development of the EDTA quench method. This work was supported by Academy of Finland Grant 286205 to G.A.B., NIH grants R01 GM087350 and R35 GM131860 to K.S.M. and Sigrid Jusélius Foundation grant 1702 to M.M-K. and G.A.B.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - RNA polymerases (RNAPs) synthesize RNA from NTPs, whereas DNA polymerases synthesize DNA from 2′dNTPs. DNA polymerases select against NTPs by using steric gates to exclude the 2′OH, but RNAPs have to employ alternative selection strategies. In single-subunit RNAPs, a conserved Tyr residue discriminates against 2′dNTPs, whereas selectivity mechanisms of multi-subunit RNAPs remain hitherto unknown. Here, we show that a conserved Arg residue uses a two-pronged strategy to select against 2′dNTPs in multi-subunit RNAPs. The conserved Arg interacts with the 2′OH group to promote NTP binding, but selectively inhibits incorporation of 2′dNTPs by interacting with their 3′OH group to favor the catalytically-inert 2′-endo conformation of the deoxyribose moiety. This deformative action is an elegant example of an active selection against a substrate that is a substructure of the correct substrate. Our findings provide important insights into the evolutionary origins of biopolymers and the design of selective inhibitors of viral RNAPs.
AB - RNA polymerases (RNAPs) synthesize RNA from NTPs, whereas DNA polymerases synthesize DNA from 2′dNTPs. DNA polymerases select against NTPs by using steric gates to exclude the 2′OH, but RNAPs have to employ alternative selection strategies. In single-subunit RNAPs, a conserved Tyr residue discriminates against 2′dNTPs, whereas selectivity mechanisms of multi-subunit RNAPs remain hitherto unknown. Here, we show that a conserved Arg residue uses a two-pronged strategy to select against 2′dNTPs in multi-subunit RNAPs. The conserved Arg interacts with the 2′OH group to promote NTP binding, but selectively inhibits incorporation of 2′dNTPs by interacting with their 3′OH group to favor the catalytically-inert 2′-endo conformation of the deoxyribose moiety. This deformative action is an elegant example of an active selection against a substrate that is a substructure of the correct substrate. Our findings provide important insights into the evolutionary origins of biopolymers and the design of selective inhibitors of viral RNAPs.
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U2 - 10.1038/s41467-021-21005-w
DO - 10.1038/s41467-021-21005-w
M3 - Article
C2 - 33542236
AN - SCOPUS:85100449652
SN - 2041-1723
VL - 12
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 796
ER -