结核分枝杆菌PhoP-PhoR双组分系统研究进展
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摘要
双组分信号转导系统广泛存在于各种原核生物中,其基本结构为1个组氨酸激酶(HK)和1个反应调节剂(RR),该系统能够感知外界刺激并作出反应,使细菌能适应各种不利环境且能增强细菌在宿主体内的生存能力,对细菌的毒力和生长至关重要。文章概述了结核分枝杆菌DosS/DosT-DosR、MprA-MprB、PrrA-PrrB、PhoPPhoR等12个双组分系统,重点介绍了PhoP-PhoR双组分系统的结构与功能。PhoP-PhoR是结核分枝杆菌最基本、最重要的双组分系统之一,由感受器PhoR和效应器PhoP组成,其中PhoR可以接受Mg2+、Cl-或H+等外界刺激,进而驱动PhoP对靶基因的转录表达进行调控。PhoP作为一种效应蛋白,经过晶体结构解析发现,其可通过与DNA结合来实现对靶基因的调控,具体包括对细胞壁组成的控制、对脂类代谢和pH的调节、对结核分枝杆菌毒力网络的调控。此外,文章还介绍了PhoP突变株作为疫苗候选株所具有的优势,包括PhoP突变株在小鼠模型中毒力减弱,并且免疫恒河猴及豚鼠后均有一定的免疫保护性等,表明PhoP突变株具有较好的疫苗开发潜力。作者通过对PhoP-PhoR双组分系统的结构与功能,以及PhoP突变株作为疫苗候选株的研究进行综述,旨在为结核分枝杆菌的毒力机制和人类结核病疫苗的研究提供理论依据。
The two-component signal transduction system is widely distributed in various prokaryotes.Its basic structure consist of a histidine protein kinase(HK)and a response-regulatory protein(RR).The system is essential for the virulence and growth of bacteria which can perceive various external stimuli and react,so that bacteria can adapt to various adverse conditions and enhance the survival ability in the host.In this paper,12 two-component systems of Mycobacterium tuberculosis including DosS/DosT-DosR,MprA-MprB,PrrA-PrrB and PhoP-PhoR are briefly reviewed.The structure and function of PhoP-PhoR are highlighted.PhoP-PhoR is one of the most basic and most important two component systems of Mycobacterium tuberculosis,composed of receptor PhoR and effector PhoP,in which PhoR can receive external stimuli such as Mg~(2+),Cl~- or H~+,and then drive PhoP to regulate the transcriptional expression of target genes.As an effectorprotein,PhoP can be found through the analysis of crystal structure,which can be used to regulate the target gene by combining with DNA,including the control of cell wall composition,the regulation of lipid metabolism and pH,and the regulation of the virulence network of Mycobacterium tuberculosis.In addition,the advantages of PhoP mutant strain as a vaccine candidate is also introduced in the article,including the weakening of PhoP mutant strain in the mouse model,and immune protection in immunized Ganges River monkeys and guinea pigs,indicating that the PhoP mutant has a good potential for vaccine development.In this paper,the structure and function of PhoP-PhoR two-component system and the research of PhoP mutant as vaccine candidate are reviewed to provide a theoretical basis for research of the virulence mechanism of Mycobacterium tuberculosis and human tuberculosis vaccine.
The two-component signal transduction system is widely distributed in various prokaryotes.Its basic structure consist of a histidine protein kinase(HK)and a response-regulatory protein(RR).The system is essential for the virulence and growth of bacteria which can perceive various external stimuli and react,so that bacteria can adapt to various adverse conditions and enhance the survival ability in the host.In this paper,12 two-component systems of Mycobacterium tuberculosis including DosS/DosT-DosR,MprA-MprB,PrrA-PrrB and PhoP-PhoR are briefly reviewed.The structure and function of PhoP-PhoR are highlighted.PhoP-PhoR is one of the most basic and most important two component systems of Mycobacterium tuberculosis,composed of receptor PhoR and effector PhoP,in which PhoR can receive external stimuli such as Mg~(2+),Cl~- or H~+,and then drive PhoP to regulate the transcriptional expression of target genes.As an effectorprotein,PhoP can be found through the analysis of crystal structure,which can be used to regulate the target gene by combining with DNA,including the control of cell wall composition,the regulation of lipid metabolism and pH,and the regulation of the virulence network of Mycobacterium tuberculosis.In addition,the advantages of PhoP mutant strain as a vaccine candidate is also introduced in the article,including the weakening of PhoP mutant strain in the mouse model,and immune protection in immunized Ganges River monkeys and guinea pigs,indicating that the PhoP mutant has a good potential for vaccine development.In this paper,the structure and function of PhoP-PhoR two-component system and the research of PhoP mutant as vaccine candidate are reviewed to provide a theoretical basis for research of the virulence mechanism of Mycobacterium tuberculosis and human tuberculosis vaccine.
引文
[1]ZIMPEL C K,BRANDAO P E,FILHO A F D S,et al.Complete genome sequencing of Mycobacterium bovis SP38and comparative genomics of Mycobacterium bovis and M.tuberculosis strains[J].Frontiers in Microbiology,2017,8:2389.
[2]WHO.Global tuberculosis report 2017[R].2017.
[3]BRRETT J F,HOCH J A.Two-component signal transduction as a target for microbial anti-infective therapy[J].Antimicrobial Agents&Chemotherapy,1998,42(7):1529-1536.
[4]WHITE M J,HE H,PENOSKE R M,et al.PepDparticipates in the mycobacterial stress response mediated through MprAB and SigE[J].Journal of Bacteriology,2010,192:1498-1510.
[5]WHITE M J,SAVARYN J P,BRETL D J,et al.The HtrA-like serine protease PepD interacts with and modulates the Mycobacterium tuberculosis 35-kDa antigen outer envelope protein[J].PLoS One,2011,6:e18175.
[6]EWANN F,JACKSON M,PETHE K,et al.Transient requirement of the PrrA-PrrB two-component system for early intracellular multiplication of Mycobacterium tuberculosis[J].Infection and Immunity,2002,70(5):2256-2263.
[7]EWANN F,LOCHT C,SUPPLY P.Intracellular autoregulation of the Mycobacterium tuberculosis PrrAresponse regulator[J].Journal of Microbiology,2004,150:241-246.
[8]GONZALO-ASENSIO J,MOSTOWY S,HARDERS-WESTERVEEN J,et al.PhoP:A missing piece in the intricate puzzle of Mycobacterium tuberculosis virulence[J].PLoS One,2008,3(10):e3496.
[9]RIFAT D,BISHAI W R,KARAKOUSIS P C.Phosphate depletion:A novel trigger for Mycobacterium tuberculosis persistence[J].Journal of Infectious Diseases,2009,200(7):1126-1135.
[10]LEE H N,JUNG K E,KO I J,et al.Protein-protein interactions between histidine kinases and response regulators of Mycobacterium tuberculosis H37Rv[J].Journal of Microbiology,2012,50(2):270-277.
[11]CANGELOSI G A,DO J S,FREEMAN R,et al.The two-component regulatory system mtrAB is required for morphotypic multidrug resistance in Mycobacterium avium[J].Antimicrob Agents Chemother,2006,50(2):461-468.
[12]GUPTA S,SINHA A,SARKAR D.Transcriptional autoregulation by Mycobacterium tuberculosis PhoPinvolves recognition of novel direct repeat sequences in the regulatory region of the promoter[J].FEBSLetters,2006,580(22):5328-5338.
[13]GONZALO-ASENSIO J,SOTO C Y,ARBUES A,et al.The Mycobacterium tuberculosis phoPR operon is positively autoregulated in the virulent strain H37Rv[J].Journal of Bacteriology,2008,190:7068-7078.
[14]RAGHAVAN S,P MANZANILLO,CHAN K,et al.Secreted transcription factor controls Mycobacterium tuberculosis virulence[J].Nature,2008,454(7205):717-721.
[15]BAKER J J,JOHNSON B K,ABRAMOVITCH R B.Slow growth of Mycobacterium tuberculosis at acidic pH is regulated by phoPR and host-associated carbon sources[J].Molecular Microbiology,2014,94(1):56-69.
[16]VIDYARTHI A,KHAN N,AGNIHOTRI T,et al.Antibody response against PhoP efficiently discriminates among healthy individuals,tuberculosis patients and their contacts[J].PLoS One,2017,12(3):e0173769.
[17]WANG L,MIAO X,SOUTHALL N,et al.A highthroughput assay for developing inhibitors of PhoP,a virulence factor of Mycobacterium tuberculosis[J].Combinatorial Chemistry&High Throughput Screening,2016,19(10):855-864.
[18]PATHAK A,GOYAL R,SINHA A,et al.Domain structure of virulence-associated response regulator PhoP of Mycobacterium tuberculosis:Role of the linker region in regulator-promoter interaction(s)[J].Journal of Biological Chemistry,2010,285(45):34309-34318.
[19]MENON S,WANG S.Structure of the response regulator PhoP from Mycobacterium tuberculosis reveals a dimer through the receiver domain[J].Biochemistry,2011,50(26):5948-5957.
[20]MACDONALD R,SRAKAR D,AMER B R,et al.Solution structure of the PhoP DNA-binding domain from Mycobacterium tuberculosis[J].Journal of Biomolecular NMR,2015,63(1):111-117.
[21]WANG S S,JENA E,ISSAR S.Structure of the DNA-binding domain of the response regulator PhoPfrom Mycobacterium tuberculosis[J].Biochemistry,2007,46(51):14751-14761.
[22]CHESNE-SECK M L,BARILONE N,BOUDUN F,et al.A point mutation in the two-component regulator PhoP-PhoR accounts for the absence of polyketide-derived acyltrehaloses but not that of phthiocerol dimycocerosates in Mycobacterium tuberculosis H37Ra[J].Journal of Bacteriology,2008,190(4):1329-1334.
[23]GOYAL R,DAS A K,SINGH R,et al.Phosphorylation of PhoP protein plays a direct regulatory role in lipid biosynthesis of Mycobacterium tuberculosis[J].Journal of Biological Chemistry,2011,286(52):45197-45208.
[24]ZENG J,CUI T,HE Z G.A genome-wide regulatorDNA interaction network in the human pathogen Mycobacterium tuberculosis H37Rv[J].Journal of Proteome Research,2012,11(9):4682-4692.
[25]SINHA A,GUPTA S,BHUTANI S,et al.PhoP-PhoP interaction at adjacent PhoP binding sites is influenced by protein phosphorylation[J].Journal of Bacteriology,2008,190(4):1317-1328.
[26]HE X,WANG S.DNA consensus sequence motif for binding response regulator PhoP,a virulence regulator of Mycobacterium tuberculosis[J].Biochemistry,2014,53(51):8008-8020.
[27]CIMINO M,THOMAS C,NAMOUCHI A,et al.Identification of DNA binding motifs of the Mycobacterium tuberculosis PhoP/PhoR two-component signal transduction system[J].PLoS One,2012,7(8):e42876.
[28]HE X,WANG L,WANG S.Structural basis of DNAsequence recognition by the response regulator PhoPin Mycobacterium tuberculosis[J].Scientific Reports,2016,6(a1):24442.
[29]LUDWICZAK P,GILLERON M,BORDAT Y,et al.Mycobacterium tuberculosis phoP mutant:Lipoarabinomannan molecular structure[J].Microbiology,2002,148(10):3029-3037.
[30]GOUDE R,AMIN A G,CHATTERJEE D,et al.The critical role of embC in Mycobacterium tuberculosis[J].Journal of Bacteriology,2008,190(12):4335-4341.
[31]GOUDE R,PARISH T.The genetics of cell wall biosynthesis in Mycobacterium tuberculosis[J].Future Microbiol,2008,3(3):299-313.
[32]BRENNAN P J.Structure,function,and biogenesis of the cell wall of Mycobacterium tuberculosis[J].Tuberculosis,2003,83:91-97.
[33]WALTERS S B,DUBNAU E,KOLESNIKOVA I,et al.The Mycobacterium tuberculosis PhoPR twocomponent system regulates genes essential for virulence and complex lipid biosynthesis[J].Molecular Microbiology,2006,60(2):312-330.
[34]ASENSIO J G,MAIA C,FERRER N L,et al.The virulence associated two component PhoP-PhoR system controls the biosynthesis of polyketide-derived lipids in Mycobacterium tuberculosis[J].Journal of Biological Chemistry,2006,281(3):1313-1316.
[35]LI A H,WADDELL S J,HINDS J,et al.Contrasting transcriptional responses of a virulent and an attenuated strain of Mycobacterium tuberculosis infecting macrophages[J].PLoS One,2010,5(6):e11066.
[36]ABRAMOVITCH R B,ROHDE K H,FONG F H,et al.AprABC:A Mycobacterium tuberculosis complex-specific locus that modulates pH-driven adaptation to the macrophage phagosome[J].Molecular Microbiology,2011,80(3):678-694.
[37]JESUS G A,SERGE M,JOSE H W,et al.PhoP:Amissing piece in the intricate puzzle of Mycobacterium tuberculosis virulence[J].PLoS One,2008,3(10):e3496.
[38]KUMAR V A,GOYAL R,BANSAL R,et al.EspR-dependent ESAT-6protein secretion of Mycobacterium tuberculosis requires the presence of virulence regulator PhoP[J].Journal of Biological Chemistry,2016,291(36):19018-19030.
[39]JOHNSON B K,COLVIN C J,NEEDLE D B,et al.The carbonic anhydrase inhibitor ethoxzolamide inhibits the Mycobacterium tuberculosis PhoPR Regulon and Esx-1secretion and attenuates virulence[J].Antimicrobial Agents&Chemotherapy,2015,59(8):4436-4445.
[40]MARTIN C,WILLIAMS A,HERNANDZE-PANDO R,et al.The live Mycobacterium tuberculosis phoP,mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs[J].Vaccine,2006,24(17):3408-3419.
[41]VERRECK F A W,VERVENNE R A W,KONDOVA I,et al.MVA.85Aboosting of BCG and an attenuated,phoP deficient Mycobacterium tuberculosis vaccine both show protective efficacy against tuberculosis in rhesus macaques[J].PLoS One,2009,4(4):e5264.
[42]CARDONA P J,ASENSIO J G,ARBUES A,et al.Extended safety studies of the attenuated live tuberculosis vaccine SO2 based on phoP mutant[J].Vaccine,2009,27(18):2499-2505.
[43]ARBUES A,AGUILO J I,GONZALO-ASENSIO J,et al.Construction characterization and preclinical evaluation of MTBVAC,the first live-attenuated Mycobacterium tuberculosis-based vaccine to enter clinical trials[J].Vaccine,2013,31(42):4867-4873.
[2]WHO.Global tuberculosis report 2017[R].2017.
[3]BRRETT J F,HOCH J A.Two-component signal transduction as a target for microbial anti-infective therapy[J].Antimicrobial Agents&Chemotherapy,1998,42(7):1529-1536.
[4]WHITE M J,HE H,PENOSKE R M,et al.PepDparticipates in the mycobacterial stress response mediated through MprAB and SigE[J].Journal of Bacteriology,2010,192:1498-1510.
[5]WHITE M J,SAVARYN J P,BRETL D J,et al.The HtrA-like serine protease PepD interacts with and modulates the Mycobacterium tuberculosis 35-kDa antigen outer envelope protein[J].PLoS One,2011,6:e18175.
[6]EWANN F,JACKSON M,PETHE K,et al.Transient requirement of the PrrA-PrrB two-component system for early intracellular multiplication of Mycobacterium tuberculosis[J].Infection and Immunity,2002,70(5):2256-2263.
[7]EWANN F,LOCHT C,SUPPLY P.Intracellular autoregulation of the Mycobacterium tuberculosis PrrAresponse regulator[J].Journal of Microbiology,2004,150:241-246.
[8]GONZALO-ASENSIO J,MOSTOWY S,HARDERS-WESTERVEEN J,et al.PhoP:A missing piece in the intricate puzzle of Mycobacterium tuberculosis virulence[J].PLoS One,2008,3(10):e3496.
[9]RIFAT D,BISHAI W R,KARAKOUSIS P C.Phosphate depletion:A novel trigger for Mycobacterium tuberculosis persistence[J].Journal of Infectious Diseases,2009,200(7):1126-1135.
[10]LEE H N,JUNG K E,KO I J,et al.Protein-protein interactions between histidine kinases and response regulators of Mycobacterium tuberculosis H37Rv[J].Journal of Microbiology,2012,50(2):270-277.
[11]CANGELOSI G A,DO J S,FREEMAN R,et al.The two-component regulatory system mtrAB is required for morphotypic multidrug resistance in Mycobacterium avium[J].Antimicrob Agents Chemother,2006,50(2):461-468.
[12]GUPTA S,SINHA A,SARKAR D.Transcriptional autoregulation by Mycobacterium tuberculosis PhoPinvolves recognition of novel direct repeat sequences in the regulatory region of the promoter[J].FEBSLetters,2006,580(22):5328-5338.
[13]GONZALO-ASENSIO J,SOTO C Y,ARBUES A,et al.The Mycobacterium tuberculosis phoPR operon is positively autoregulated in the virulent strain H37Rv[J].Journal of Bacteriology,2008,190:7068-7078.
[14]RAGHAVAN S,P MANZANILLO,CHAN K,et al.Secreted transcription factor controls Mycobacterium tuberculosis virulence[J].Nature,2008,454(7205):717-721.
[15]BAKER J J,JOHNSON B K,ABRAMOVITCH R B.Slow growth of Mycobacterium tuberculosis at acidic pH is regulated by phoPR and host-associated carbon sources[J].Molecular Microbiology,2014,94(1):56-69.
[16]VIDYARTHI A,KHAN N,AGNIHOTRI T,et al.Antibody response against PhoP efficiently discriminates among healthy individuals,tuberculosis patients and their contacts[J].PLoS One,2017,12(3):e0173769.
[17]WANG L,MIAO X,SOUTHALL N,et al.A highthroughput assay for developing inhibitors of PhoP,a virulence factor of Mycobacterium tuberculosis[J].Combinatorial Chemistry&High Throughput Screening,2016,19(10):855-864.
[18]PATHAK A,GOYAL R,SINHA A,et al.Domain structure of virulence-associated response regulator PhoP of Mycobacterium tuberculosis:Role of the linker region in regulator-promoter interaction(s)[J].Journal of Biological Chemistry,2010,285(45):34309-34318.
[19]MENON S,WANG S.Structure of the response regulator PhoP from Mycobacterium tuberculosis reveals a dimer through the receiver domain[J].Biochemistry,2011,50(26):5948-5957.
[20]MACDONALD R,SRAKAR D,AMER B R,et al.Solution structure of the PhoP DNA-binding domain from Mycobacterium tuberculosis[J].Journal of Biomolecular NMR,2015,63(1):111-117.
[21]WANG S S,JENA E,ISSAR S.Structure of the DNA-binding domain of the response regulator PhoPfrom Mycobacterium tuberculosis[J].Biochemistry,2007,46(51):14751-14761.
[22]CHESNE-SECK M L,BARILONE N,BOUDUN F,et al.A point mutation in the two-component regulator PhoP-PhoR accounts for the absence of polyketide-derived acyltrehaloses but not that of phthiocerol dimycocerosates in Mycobacterium tuberculosis H37Ra[J].Journal of Bacteriology,2008,190(4):1329-1334.
[23]GOYAL R,DAS A K,SINGH R,et al.Phosphorylation of PhoP protein plays a direct regulatory role in lipid biosynthesis of Mycobacterium tuberculosis[J].Journal of Biological Chemistry,2011,286(52):45197-45208.
[24]ZENG J,CUI T,HE Z G.A genome-wide regulatorDNA interaction network in the human pathogen Mycobacterium tuberculosis H37Rv[J].Journal of Proteome Research,2012,11(9):4682-4692.
[25]SINHA A,GUPTA S,BHUTANI S,et al.PhoP-PhoP interaction at adjacent PhoP binding sites is influenced by protein phosphorylation[J].Journal of Bacteriology,2008,190(4):1317-1328.
[26]HE X,WANG S.DNA consensus sequence motif for binding response regulator PhoP,a virulence regulator of Mycobacterium tuberculosis[J].Biochemistry,2014,53(51):8008-8020.
[27]CIMINO M,THOMAS C,NAMOUCHI A,et al.Identification of DNA binding motifs of the Mycobacterium tuberculosis PhoP/PhoR two-component signal transduction system[J].PLoS One,2012,7(8):e42876.
[28]HE X,WANG L,WANG S.Structural basis of DNAsequence recognition by the response regulator PhoPin Mycobacterium tuberculosis[J].Scientific Reports,2016,6(a1):24442.
[29]LUDWICZAK P,GILLERON M,BORDAT Y,et al.Mycobacterium tuberculosis phoP mutant:Lipoarabinomannan molecular structure[J].Microbiology,2002,148(10):3029-3037.
[30]GOUDE R,AMIN A G,CHATTERJEE D,et al.The critical role of embC in Mycobacterium tuberculosis[J].Journal of Bacteriology,2008,190(12):4335-4341.
[31]GOUDE R,PARISH T.The genetics of cell wall biosynthesis in Mycobacterium tuberculosis[J].Future Microbiol,2008,3(3):299-313.
[32]BRENNAN P J.Structure,function,and biogenesis of the cell wall of Mycobacterium tuberculosis[J].Tuberculosis,2003,83:91-97.
[33]WALTERS S B,DUBNAU E,KOLESNIKOVA I,et al.The Mycobacterium tuberculosis PhoPR twocomponent system regulates genes essential for virulence and complex lipid biosynthesis[J].Molecular Microbiology,2006,60(2):312-330.
[34]ASENSIO J G,MAIA C,FERRER N L,et al.The virulence associated two component PhoP-PhoR system controls the biosynthesis of polyketide-derived lipids in Mycobacterium tuberculosis[J].Journal of Biological Chemistry,2006,281(3):1313-1316.
[35]LI A H,WADDELL S J,HINDS J,et al.Contrasting transcriptional responses of a virulent and an attenuated strain of Mycobacterium tuberculosis infecting macrophages[J].PLoS One,2010,5(6):e11066.
[36]ABRAMOVITCH R B,ROHDE K H,FONG F H,et al.AprABC:A Mycobacterium tuberculosis complex-specific locus that modulates pH-driven adaptation to the macrophage phagosome[J].Molecular Microbiology,2011,80(3):678-694.
[37]JESUS G A,SERGE M,JOSE H W,et al.PhoP:Amissing piece in the intricate puzzle of Mycobacterium tuberculosis virulence[J].PLoS One,2008,3(10):e3496.
[38]KUMAR V A,GOYAL R,BANSAL R,et al.EspR-dependent ESAT-6protein secretion of Mycobacterium tuberculosis requires the presence of virulence regulator PhoP[J].Journal of Biological Chemistry,2016,291(36):19018-19030.
[39]JOHNSON B K,COLVIN C J,NEEDLE D B,et al.The carbonic anhydrase inhibitor ethoxzolamide inhibits the Mycobacterium tuberculosis PhoPR Regulon and Esx-1secretion and attenuates virulence[J].Antimicrobial Agents&Chemotherapy,2015,59(8):4436-4445.
[40]MARTIN C,WILLIAMS A,HERNANDZE-PANDO R,et al.The live Mycobacterium tuberculosis phoP,mutant strain is more attenuated than BCG and confers protective immunity against tuberculosis in mice and guinea pigs[J].Vaccine,2006,24(17):3408-3419.
[41]VERRECK F A W,VERVENNE R A W,KONDOVA I,et al.MVA.85Aboosting of BCG and an attenuated,phoP deficient Mycobacterium tuberculosis vaccine both show protective efficacy against tuberculosis in rhesus macaques[J].PLoS One,2009,4(4):e5264.
[42]CARDONA P J,ASENSIO J G,ARBUES A,et al.Extended safety studies of the attenuated live tuberculosis vaccine SO2 based on phoP mutant[J].Vaccine,2009,27(18):2499-2505.
[43]ARBUES A,AGUILO J I,GONZALO-ASENSIO J,et al.Construction characterization and preclinical evaluation of MTBVAC,the first live-attenuated Mycobacterium tuberculosis-based vaccine to enter clinical trials[J].Vaccine,2013,31(42):4867-4873.