TY - JOUR
T1 - Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides
AU - Tchigvintsev, Anatoli
AU - Tchigvintsev, Dmitri
AU - Flick, Robert
AU - Popovic, Ana
AU - Dong, Aiping
AU - Xu, Xiaohui
AU - Brown, Greg
AU - Lu, Wenyun
AU - Wu, Hong
AU - Cui, Hong
AU - Dombrowski, Ludmila
AU - Joo, Jeong Chan
AU - Beloglazova, Natalia
AU - Min, Jinrong
AU - Savchenko, Alexei
AU - Caudy, Amy A.
AU - Rabinowitz, Joshua D.
AU - Murzin, Alexey G.
AU - Yakunin, Alexander F.
N1 - Funding Information:
This work was supported in part by the Government of Canada through Genome Canada and the Ontario Genomics Institute (2009-OGI-ABC-1405 and the research agreement OGI-055), the Ontario Research Fund (ORF-GL2-01-004 to A.F.Y.), a COMBREX grant (to A.F.Y.), the UK Medical Research Council (MC_U105192716 to A.G.M.), the Natural Sciences and Engineering Research Council and Canadian Institutes of Health Research (to A.A.C.), and National Institutes of Health grants P50GM071508 and 1R01CA16359-01A1 (to J.D.R.). The Structural Genomics Consortium is a registered charity (number 1097737) that receives funds from the Canada Foundation for Innovation, Eli Lilly Canada, GlaxoSmithKline, the Ontario Ministry of Economic Development and Innovation, the Novartis Research Foundation, Pfizer, AbbVie, Takeda, Janssen, Boehringer Ingelheim, and the Wellcome Trust.
PY - 2013/11/21
Y1 - 2013/11/21
N2 - Maf (for multicopy associated filamentation) proteins represent a large family of conserved proteins implicated in cell division arrest but whose biochemical activity remains unknown. Here, we show that the prokaryotic and eukaryotic Maf proteins exhibit nucleotide pyrophosphatase activity against 5-methyl-UTP, pseudo-UTP, 5-methyl-CTP, and 7-methyl-GTP, which represent the most abundant modified bases in all organisms, as well as against canonical nucleotides dTTP, UTP, and CTP. Overexpression of the Maf protein YhdE in E. coli cells increased intracellular levels of dTMP and UMP, confirming that dTTP and UTP are the in vivo substrates of this protein. Crystal structures and site-directed mutagenesis of Maf proteins revealed the determinants of their activity and substrate specificity. Thus, pyrophosphatase activity of Maf proteins toward canonical and modified nucleotides might provide the molecular mechanism for a dual role of these proteins in cell division arrest and house cleaning.
AB - Maf (for multicopy associated filamentation) proteins represent a large family of conserved proteins implicated in cell division arrest but whose biochemical activity remains unknown. Here, we show that the prokaryotic and eukaryotic Maf proteins exhibit nucleotide pyrophosphatase activity against 5-methyl-UTP, pseudo-UTP, 5-methyl-CTP, and 7-methyl-GTP, which represent the most abundant modified bases in all organisms, as well as against canonical nucleotides dTTP, UTP, and CTP. Overexpression of the Maf protein YhdE in E. coli cells increased intracellular levels of dTMP and UMP, confirming that dTTP and UTP are the in vivo substrates of this protein. Crystal structures and site-directed mutagenesis of Maf proteins revealed the determinants of their activity and substrate specificity. Thus, pyrophosphatase activity of Maf proteins toward canonical and modified nucleotides might provide the molecular mechanism for a dual role of these proteins in cell division arrest and house cleaning.
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U2 - 10.1016/j.chembiol.2013.09.011
DO - 10.1016/j.chembiol.2013.09.011
M3 - Article
C2 - 24210219
AN - SCOPUS:84888296777
SN - 1074-5521
VL - 20
SP - 1386
EP - 1398
JO - Chemistry and Biology
JF - Chemistry and Biology
IS - 11
ER -