Structural basis of response regulator dephosphorylation by Rap phosphatases.

Bacterial Rap family proteins have been most extensively studied in Bacillus subtilis, where they regulate activities including sporulation, genetic competence, antibiotic expression, and the movement of the ICEBs1 transposon. One subset of Rap proteins consists of phosphatases that control B. subti...

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Autores principales:	Vijay Parashar, Nicolas Mirouze, David A Dubnau, Matthew B Neiditch
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/646ca44ec642421aa25cb8259ee3ebae
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spelling	oai:doaj.org-article:646ca44ec642421aa25cb8259ee3ebae2021-11-18T05:36:20ZStructural basis of response regulator dephosphorylation by Rap phosphatases.1544-91731545-788510.1371/journal.pbio.1000589https://doaj.org/article/646ca44ec642421aa25cb8259ee3ebae2011-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21346797/?tool=EBIhttps://doaj.org/toc/1544-9173https://doaj.org/toc/1545-7885Bacterial Rap family proteins have been most extensively studied in Bacillus subtilis, where they regulate activities including sporulation, genetic competence, antibiotic expression, and the movement of the ICEBs1 transposon. One subset of Rap proteins consists of phosphatases that control B. subtilis and B. anthracis sporulation by dephosphorylating the response regulator Spo0F. The mechanistic basis of Rap phosphatase activity was unknown. Here we present the RapH-Spo0F X-ray crystal structure, which shows that Rap proteins consist of a 3-helix bundle and a tetratricopeptide repeat domain. Extensive biochemical and genetic functional studies reveal the importance of the observed RapH-Spo0F interactions, including the catalytic role of a glutamine in the RapH 3-helix bundle that inserts into the Spo0F active site. We show that in addition to dephosphorylating Spo0F, RapH can antagonize sporulation by sterically blocking phosphoryl transfer to and from Spo0F. Our structure-function analysis of the RapH-Spo0F interaction identified Rap protein residues critical for Spo0F phosphatase activity. This information enabled us to assign Spo0F phosphatase activity to a Rap protein based on sequence alone, which was not previously possible. Finally, as the ultimate test of our newfound understanding of the structural requirements for Rap phosphatase function, a non-phosphatase Rap protein that inhibits the binding of the response regulator ComA to DNA was rationally engineered to dephosphorylate Spo0F. In addition to revealing the mechanistic basis of response regulator dephosphorylation by Rap proteins, our studies support the previously proposed T-loop-Y allostery model of receiver domain regulation that restricts the aromatic "switch" residue to an internal position when the β4-α4 loop adopts an active-site proximal conformation.Vijay ParasharNicolas MirouzeDavid A DubnauMatthew B NeiditchPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Biology, Vol 9, Iss 2, p e1000589 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Vijay Parashar Nicolas Mirouze David A Dubnau Matthew B Neiditch Structural basis of response regulator dephosphorylation by Rap phosphatases.
description	Bacterial Rap family proteins have been most extensively studied in Bacillus subtilis, where they regulate activities including sporulation, genetic competence, antibiotic expression, and the movement of the ICEBs1 transposon. One subset of Rap proteins consists of phosphatases that control B. subtilis and B. anthracis sporulation by dephosphorylating the response regulator Spo0F. The mechanistic basis of Rap phosphatase activity was unknown. Here we present the RapH-Spo0F X-ray crystal structure, which shows that Rap proteins consist of a 3-helix bundle and a tetratricopeptide repeat domain. Extensive biochemical and genetic functional studies reveal the importance of the observed RapH-Spo0F interactions, including the catalytic role of a glutamine in the RapH 3-helix bundle that inserts into the Spo0F active site. We show that in addition to dephosphorylating Spo0F, RapH can antagonize sporulation by sterically blocking phosphoryl transfer to and from Spo0F. Our structure-function analysis of the RapH-Spo0F interaction identified Rap protein residues critical for Spo0F phosphatase activity. This information enabled us to assign Spo0F phosphatase activity to a Rap protein based on sequence alone, which was not previously possible. Finally, as the ultimate test of our newfound understanding of the structural requirements for Rap phosphatase function, a non-phosphatase Rap protein that inhibits the binding of the response regulator ComA to DNA was rationally engineered to dephosphorylate Spo0F. In addition to revealing the mechanistic basis of response regulator dephosphorylation by Rap proteins, our studies support the previously proposed T-loop-Y allostery model of receiver domain regulation that restricts the aromatic "switch" residue to an internal position when the β4-α4 loop adopts an active-site proximal conformation.
format	article
author	Vijay Parashar Nicolas Mirouze David A Dubnau Matthew B Neiditch
author_facet	Vijay Parashar Nicolas Mirouze David A Dubnau Matthew B Neiditch
author_sort	Vijay Parashar
title	Structural basis of response regulator dephosphorylation by Rap phosphatases.
title_short	Structural basis of response regulator dephosphorylation by Rap phosphatases.
title_full	Structural basis of response regulator dephosphorylation by Rap phosphatases.
title_fullStr	Structural basis of response regulator dephosphorylation by Rap phosphatases.
title_full_unstemmed	Structural basis of response regulator dephosphorylation by Rap phosphatases.
title_sort	structural basis of response regulator dephosphorylation by rap phosphatases.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/646ca44ec642421aa25cb8259ee3ebae
work_keys_str_mv	AT vijayparashar structuralbasisofresponseregulatordephosphorylationbyrapphosphatases AT nicolasmirouze structuralbasisofresponseregulatordephosphorylationbyrapphosphatases AT davidadubnau structuralbasisofresponseregulatordephosphorylationbyrapphosphatases AT matthewbneiditch structuralbasisofresponseregulatordephosphorylationbyrapphosphatases
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Structural basis of response regulator dephosphorylation by Rap phosphatases.

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