Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides

Anatoli Tchigvintsev; Dmitri Tchigvintsev; Robert Flick; Ana Popovic; Aiping Dong; Xiaohui Xu; Greg Brown; Wenyun Lu; Hong Wu; Hong Cui; Ludmila Dombrowski; Jeong Chan Joo; Natalia Beloglazova; Jinrong Min; Alexei Savchenko; Amy A. Caudy; Joshua D. Rabinowitz; Alexey G. Murzin; Alexander F. Yakunin

doi:10.1016/j.chembiol.2013.09.011

Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides

Anatoli Tchigvintsev, Dmitri Tchigvintsev, Robert Flick, Ana Popovic, Aiping Dong, Xiaohui Xu, Greg Brown, Wenyun Lu, Hong Wu, Hong Cui, Ludmila Dombrowski, Jeong Chan Joo, Natalia Beloglazova, Jinrong Min, Alexei Savchenko, Amy A. Caudy, Joshua D. Rabinowitz, Alexey G. Murzin, Alexander F. Yakunin

Department of Biotechnology

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	1386-1398
Number of pages	13
Journal	Chemistry and Biology
Volume	20
Issue number	11
DOIs	https://doi.org/10.1016/j.chembiol.2013.09.011
State	Published - 21 Nov 2013

Bibliographical note

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.

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Tchigvintsev, A., Tchigvintsev, D., Flick, R., Popovic, A., Dong, A., Xu, X., Brown, G., Lu, W., Wu, H., Cui, H., Dombrowski, L., Joo, J. C., Beloglazova, N., Min, J., Savchenko, A., Caudy, A. A., Rabinowitz, J. D., Murzin, A. G., & Yakunin, A. F. (2013). Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides. Chemistry and Biology, 20(11), 1386-1398. https://doi.org/10.1016/j.chembiol.2013.09.011

@article{323f36ba9e074ca69ec2767c3dfaf053,

title = "Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides",

abstract = "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.",

author = "Anatoli Tchigvintsev and Dmitri Tchigvintsev and Robert Flick and Ana Popovic and Aiping Dong and Xiaohui Xu and Greg Brown and Wenyun Lu and Hong Wu and Hong Cui and Ludmila Dombrowski and Joo, {Jeong Chan} and Natalia Beloglazova and Jinrong Min and Alexei Savchenko and Caudy, {Amy A.} and Rabinowitz, {Joshua D.} and Murzin, {Alexey G.} and Yakunin, {Alexander F.}",

note = "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. ",

year = "2013",

month = nov,

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doi = "10.1016/j.chembiol.2013.09.011",

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Tchigvintsev, A, Tchigvintsev, D, Flick, R, Popovic, A, Dong, A, Xu, X, Brown, G, Lu, W, Wu, H, Cui, H, Dombrowski, L, Joo, JC, Beloglazova, N, Min, J, Savchenko, A, Caudy, AA, Rabinowitz, JD, Murzin, AG & Yakunin, AF 2013, 'Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides', Chemistry and Biology, vol. 20, no. 11, pp. 1386-1398. https://doi.org/10.1016/j.chembiol.2013.09.011

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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DO - 10.1016/j.chembiol.2013.09.011

M3 - Article

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AN - SCOPUS:84888296777

SN - 1074-5521

VL - 20

SP - 1386

EP - 1398

JO - Chemistry and Biology

JF - Chemistry and Biology

IS - 11

ER -

Biochemical and structural studies of conserved Maf proteins revealed nucleotide pyrophosphatases with a preference for modified nucleotides

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