Chathurange B. Ranaweera , Sunitha Shiva , Swetha Madesh , Deepika Chauhan , Roman R. Ganta , Michal Zolkiewski
{"title":"噬噬原虫 ClpB 和 DnaK 的生化特征。","authors":"Chathurange B. Ranaweera , Sunitha Shiva , Swetha Madesh , Deepika Chauhan , Roman R. Ganta , Michal Zolkiewski","doi":"10.1016/j.cstres.2024.06.003","DOIUrl":null,"url":null,"abstract":"<div><p><em>Anaplasma phagocytophilum</em> is an intracellular tick-transmitted bacterial pathogen that infects neutrophils in mammals and causes granulocytic anaplasmosis. In this study, we investigated the molecular chaperones ClpB and DnaK from <em>A. phagocytophilum</em>. In <em>Escherichia coli</em>, ClpB cooperates with DnaK and its co-chaperones DnaJ and GrpE in ATP-dependent reactivation of aggregated proteins. Since ClpB is not produced in metazoans, it is a promising target for developing antimicrobial therapies, which generates interest in studies on that chaperone’s role in pathogenic bacteria. We found that ClpB and DnaK are transcriptionally upregulated in <em>A. phagocytophilum</em> 3–5 days after infection of human HL-60 and tick ISE6 cells, which suggests an essential role of the chaperones in supporting the pathogen’s intracellular life cycle. Multiple sequence alignments show that <em>A. phagocytophilum</em> ClpB and DnaK contain all structural domains that were identified in their previously studied orthologs from other bacteria. Both <em>A. phagocytophilum</em> ClpB and DnaK display ATPase activity, which is consistent with their participation in the ATP-dependent protein disaggregation system. However, despite a significant sequence similarity between the chaperones from <em>A. phagocytophilum</em> and those from <em>E. coli</em>, the former were not as effective as their <em>E. coli</em> orthologs during reactivation of aggregated proteins <em>in vitro</em> and in supporting the survival of <em>E. coli</em> cells under heat stress. We conclude that the <em>A. phagocytophilum</em> chaperones might have evolved with distinct biochemical properties to maintain the integrity of pathogenic proteins under unique stress conditions of an intracellular environment of host cells.</p></div>","PeriodicalId":9684,"journal":{"name":"Cell Stress & Chaperones","volume":"29 4","pages":"Pages 540-551"},"PeriodicalIF":3.3000,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1355814524001068/pdfft?md5=49daff0ee5c7e6a52943d2057ee3c41f&pid=1-s2.0-S1355814524001068-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Biochemical characterization of ClpB and DnaK from Anaplasma phagocytophilum\",\"authors\":\"Chathurange B. Ranaweera , Sunitha Shiva , Swetha Madesh , Deepika Chauhan , Roman R. Ganta , Michal Zolkiewski\",\"doi\":\"10.1016/j.cstres.2024.06.003\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p><em>Anaplasma phagocytophilum</em> is an intracellular tick-transmitted bacterial pathogen that infects neutrophils in mammals and causes granulocytic anaplasmosis. In this study, we investigated the molecular chaperones ClpB and DnaK from <em>A. phagocytophilum</em>. In <em>Escherichia coli</em>, ClpB cooperates with DnaK and its co-chaperones DnaJ and GrpE in ATP-dependent reactivation of aggregated proteins. Since ClpB is not produced in metazoans, it is a promising target for developing antimicrobial therapies, which generates interest in studies on that chaperone’s role in pathogenic bacteria. We found that ClpB and DnaK are transcriptionally upregulated in <em>A. phagocytophilum</em> 3–5 days after infection of human HL-60 and tick ISE6 cells, which suggests an essential role of the chaperones in supporting the pathogen’s intracellular life cycle. Multiple sequence alignments show that <em>A. phagocytophilum</em> ClpB and DnaK contain all structural domains that were identified in their previously studied orthologs from other bacteria. Both <em>A. phagocytophilum</em> ClpB and DnaK display ATPase activity, which is consistent with their participation in the ATP-dependent protein disaggregation system. However, despite a significant sequence similarity between the chaperones from <em>A. phagocytophilum</em> and those from <em>E. coli</em>, the former were not as effective as their <em>E. coli</em> orthologs during reactivation of aggregated proteins <em>in vitro</em> and in supporting the survival of <em>E. coli</em> cells under heat stress. We conclude that the <em>A. phagocytophilum</em> chaperones might have evolved with distinct biochemical properties to maintain the integrity of pathogenic proteins under unique stress conditions of an intracellular environment of host cells.</p></div>\",\"PeriodicalId\":9684,\"journal\":{\"name\":\"Cell Stress & Chaperones\",\"volume\":\"29 4\",\"pages\":\"Pages 540-551\"},\"PeriodicalIF\":3.3000,\"publicationDate\":\"2024-06-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S1355814524001068/pdfft?md5=49daff0ee5c7e6a52943d2057ee3c41f&pid=1-s2.0-S1355814524001068-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cell Stress & Chaperones\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1355814524001068\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"CELL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cell Stress & Chaperones","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1355814524001068","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
Biochemical characterization of ClpB and DnaK from Anaplasma phagocytophilum
Anaplasma phagocytophilum is an intracellular tick-transmitted bacterial pathogen that infects neutrophils in mammals and causes granulocytic anaplasmosis. In this study, we investigated the molecular chaperones ClpB and DnaK from A. phagocytophilum. In Escherichia coli, ClpB cooperates with DnaK and its co-chaperones DnaJ and GrpE in ATP-dependent reactivation of aggregated proteins. Since ClpB is not produced in metazoans, it is a promising target for developing antimicrobial therapies, which generates interest in studies on that chaperone’s role in pathogenic bacteria. We found that ClpB and DnaK are transcriptionally upregulated in A. phagocytophilum 3–5 days after infection of human HL-60 and tick ISE6 cells, which suggests an essential role of the chaperones in supporting the pathogen’s intracellular life cycle. Multiple sequence alignments show that A. phagocytophilum ClpB and DnaK contain all structural domains that were identified in their previously studied orthologs from other bacteria. Both A. phagocytophilum ClpB and DnaK display ATPase activity, which is consistent with their participation in the ATP-dependent protein disaggregation system. However, despite a significant sequence similarity between the chaperones from A. phagocytophilum and those from E. coli, the former were not as effective as their E. coli orthologs during reactivation of aggregated proteins in vitro and in supporting the survival of E. coli cells under heat stress. We conclude that the A. phagocytophilum chaperones might have evolved with distinct biochemical properties to maintain the integrity of pathogenic proteins under unique stress conditions of an intracellular environment of host cells.
期刊介绍:
Cell Stress and Chaperones is an integrative journal that bridges the gap between laboratory model systems and natural populations. The journal captures the eclectic spirit of the cellular stress response field in a single, concentrated source of current information. Major emphasis is placed on the effects of climate change on individual species in the natural environment and their capacity to adapt. This emphasis expands our focus on stress biology and medicine by linking climate change effects to research on cellular stress responses of animals, micro-organisms and plants.