{"id":16763,"date":"2019-07-22T17:00:33","date_gmt":"2019-07-22T15:00:33","guid":{"rendered":"https:\/\/news.embl.de\/?p=16763"},"modified":"2024-03-22T10:56:12","modified_gmt":"2024-03-22T09:56:12","slug":"legionella-toxin","status":"publish","type":"post","link":"https:\/\/www.embl.org\/news\/science\/legionella-toxin\/","title":{"rendered":"Toxin responsible for Legionella growth identified"},"content":{"rendered":"\n

A team of scientists led by EMBL group leader Sagar Bhogaraju and Ivan Dikic of Goethe University, Frankfurt, discovered that the toxin SidJ in Legionella<\/em> bacteria enforces a unique modification on human proteins and helps legionella grow inside human cells. SidJ hijacks human protein Calmodulin to its own advantage in one of the classic examples of pathogenic bacteria exploiting the human molecular machinery and turning it against us. This makes SidJ an ideal target to curb Legionella<\/em> infection. The results have been published in Nature<\/em>.<\/p>\n\n\n\n

\"\"<\/a>
Legionella bacteria (red elliptical objects) interacting with endoplasmic reticulum. IMAGE: Goethe University<\/figcaption><\/figure><\/div>\n\n\n\n

Legionella<\/em><\/strong> \u2013 a complex bacterium<\/strong><\/h3>\n\n\n\n

Pneumonia resulting from exposure to Legionella<\/em> \u2013 although uncommon and affecting only 1 in 100,000 in Europe \u2013 has a higher than 10% fatality rate. The pathogenic bacterium Legionella pneumophila<\/em> has more than 300 toxins that it uses to infect humans. Once the aerosols containing the bacteria are inhaled, Legionella<\/em> enters the lungs where it starts infecting human cells, causing pneumonia.<\/p>\n\n\n\n

Legionella<\/em> toxins especially target the innate immune pathways facilitating the survival of the bacteria inside human cells and allowing the replication of the bacteria. Due to the large number of toxins it is difficult to see the effects of deleting one or multiple of these toxins on the Legionella<\/em> infection capacities. This is further complicated by the fact that several toxins with similar functions exist inside the bacteria. This makes Legionella<\/em> hard to target with specific drugs. <\/strong><\/p>\n\n\n\n

Focus on the SidJ toxin<\/strong><\/h3>\n\n\n\n

Researchers from EMBL Grenoble and the Goethe University in Frankfurt have now studied the toxin SidJ in detail. It is an important toxic protein of Legionella<\/em> that gets injected into the human cytoplasm and enables the successful infection and replication of the bacteria. In contrast to the other toxins in Legionella<\/em>, the deletion of SidJ alone leads to a considerable growth defect of the bacteria in human cells. This makes SidJ one of the most important toxins of Legionella<\/em> and an attractive target to curb Legionella <\/em>infection.<\/p>\n\n\n\n

While SidJ has been studied in the field for already more than a decade, the precise function of it remained unknown until today. \u201cSidJ has no sequence similarity to any of the proteins with a known function. We had to resort to standard biochemical methods and mass spectrometry to determine its function\u201d, explains Bhogaraju. \u201cWhile working out its mechanism proved to be challenging, it was also very exciting!\u201d<\/p>\n\n\n\n

In particular, the missing detailed molecular study of the toxin hindered the development of drugs that can target SidJ. The work by multidisciplinary scientists of Bhogaraju and Dikic groups now describes the molecular function of this protein in detail, elucidates its importance for Legionella<\/em> infection and provides the identity of the human proteins that are targeted by SidJ.<\/p>\n\n\n\n

Toxin at work<\/strong><\/h3>\n\n\n\n

The group showed that SidJ possesses protein glutamylation activity: it attaches the amino acid glutamate to a target protein as post-translational modification. \u201cThis kind of activity is a first for bacterial proteins\u201d, says Ivan Dikic, Director of the Institute of Biochemistry II at Goethe University. SidJ glutamylates many human proteins that are involved in tackling microbial infections and innate immunity. In order to do this, SidJ interacts with the human protein Calmodulin \u2013 a highly conserved multifunctional intermediate calcium-binding messenger protein. \u201cLegionella<\/em> has cleverly evolved to use Calmodulin to trigger SidJ’s activity and as a result prevents SidJ’s activation before the infection in the human body takes place\u201d, says Dikic.<\/p>\n\n\n\n

The cryogenic electron microscopy structure of SidJ interacting with human Calmodulin also revealed that the toxin has a kinase domain fold. “This is a both interesting and important find, as the kinase fold is druggable” says Michael Adams, a PhD student in Bhogaraju group.<\/p>\n\n\n\n

A recently study published study in Science<\/em> by Miles Black et al.<\/a> reached similar conclusions regarding the function of SidJ<\/p>\n\n\n\n

Start of a long way to therapeutic usage<\/strong><\/h3>\n\n\n\n

The outcome of the study is going to prime many studies in the future, further dissecting the mechanism of SidJ mediated glutamylation. Importantly, since the researchers found that SidJ has a kinase fold, this discovery will initiate the search for a drug molecule with potential therapeutic effects.<\/p>\n\n\n\n

\u201cWhile our work doesn\u2019t have a direct pharmaceutical application, our results on the structural and functional characteristics of one of the most important toxins of Legionella<\/em>, will lead to future studies aiming to target this protein for therapeutic uses\u201d, says Sagar Bhogaraju.<\/p>\n\n\n\n

\n\n\n\n

Toxine responsable de la croissance de Legionella<\/em> identifi\u00e9e<\/h2>\n\n\n\n

Une \u00e9quipe de scientifiques dirig\u00e9e par Sagar Bhogaraju, chef du groupe EMBL, et Ivan Dikic, de l’Universit\u00e9 Goethe de Francfort, a d\u00e9couvert que la toxine SidJ dans la bact\u00e9rie Legionella<\/em> entra\u00eene une modification unique des prot\u00e9ines humaines et favorise la croissance des l\u00e9gionelles dans les cellules humaines. SidJ d\u00e9tourne la prot\u00e9ine humaine Calmodulin \u00e0 son propre avantage dans l’un des exemples classiques de bact\u00e9ries pathog\u00e8nes exploitant la machinerie mol\u00e9culaire humaine et la retournant contre nous. Cela fait de SidJ une cible id\u00e9ale pour freiner l’infection aux l\u00e9gionelles. Les r\u00e9sultats ont \u00e9t\u00e9 publi\u00e9s dans Nature<\/em>.<\/p>\n\n\n\n

\"cryo-EM
La structure cryo-EM du complexe SidJ\/CaM. SidJ est repr\u00e9sent\u00e9 en cyan, Calmodulin en rose.<\/em>
IMAGE : EMBL<\/em><\/figcaption><\/figure><\/div>\n\n\n\n

Legionella<\/em> – une bact\u00e9rie complexe<\/strong><\/h3>\n\n\n\n

La pneumonie r\u00e9sultant d’une exposition \u00e0 la Legionella – bien qu’elle soit rare et ne touche qu’une personne sur 100 000 en Europe – a un taux de mortalit\u00e9 sup\u00e9rieur \u00e0 10%. La bact\u00e9rie pathog\u00e8ne Legionella pneumophila<\/em> poss\u00e8de plus de 300 toxines qu’elle utilise pour infecter les humains. Une fois que les a\u00e9rosols contenant la bact\u00e9rie sont inhal\u00e9s, la Legionella<\/em> p\u00e9n\u00e8tre dans les poumons o\u00f9 elle commence \u00e0 infecter les cellules humaines, provoquant une pneumonie.<\/p>\n\n\n\n

Les toxines de Legionella<\/em> ciblent particuli\u00e8rement les voies immunitaires inn\u00e9es facilitant la survie de la bact\u00e9rie \u00e0 l’int\u00e9rieur des cellules humaines et permettant la r\u00e9plication de la bact\u00e9rie. En raison du grand nombre de toxines, il est difficile de voir les effets de la suppression d’une ou de plusieurs de ces toxines sur les capacit\u00e9s d’infection \u00e0 la Legionella<\/em>. Ceci est encore compliqu\u00e9 par le fait que plusieurs toxines ayant des fonctions similaires existent \u00e0 l’int\u00e9rieur de la bact\u00e9rie. C’est pourquoi il est difficile de cibler la Legionella<\/em> avec des m\u00e9dicaments sp\u00e9cifiques.<\/p>\n\n\n\n

Zoom sur la toxine SidJ<\/strong><\/h3>\n\n\n\n

Des chercheurs de l’EMBL Grenoble et de l’Universit\u00e9 Goethe de Francfort ont maintenant \u00e9tudi\u00e9 en d\u00e9tail la toxine SidJ. C’est une prot\u00e9ine toxique importante de la Legionella<\/em> qui est inject\u00e9e dans le cytoplasme humain et qui permet l’infection et la r\u00e9plication des bact\u00e9ries. Contrairement aux autres toxines de Legionella<\/em>, la suppression de SidJ seule entra\u00eene un d\u00e9faut de croissance consid\u00e9rable de la bact\u00e9rie dans les cellules humaines. Cela fait de la SidJ l’une des toxines les plus importantes de Legionella<\/em> et une cible attrayante pour freiner l’infection \u00e0 la Legionella<\/em>.<\/p>\n\n\n\n

Bien que SidJ soit \u00e9tudi\u00e9 sur le terrain depuis d\u00e9j\u00e0 plus d’une d\u00e9cennie, sa fonction pr\u00e9cise est rest\u00e9e inconnue jusqu’\u00e0 aujourd’hui. “SidJ n’a aucune similitude de s\u00e9quence avec les prot\u00e9ines ayant une fonction connue. Nous avons d\u00fb recourir \u00e0 des m\u00e9thodes biochimiques standard et \u00e0 la spectrom\u00e9trie de masse pour d\u00e9terminer sa fonction “, explique Bhogaraju. “Si travailler sur son m\u00e9canisme s’est av\u00e9r\u00e9 \u00eatre un d\u00e9fi, c’\u00e9tait aussi tr\u00e8s excitant !”<\/p>\n\n\n\n

En particulier, l’absence d’\u00e9tude mol\u00e9culaire d\u00e9taill\u00e9e de la toxine a entrav\u00e9 le d\u00e9veloppement de m\u00e9dicaments pouvant cibler la SidJ. Les travaux de scientifiques multidisciplinaires des groupes Bhogaraju et Dikic d\u00e9crivent maintenant en d\u00e9tail la fonction mol\u00e9culaire de cette prot\u00e9ine, \u00e9lucident son importance pour l’infection \u00e0 Legionella<\/em> et fournissent l’identit\u00e9 des prot\u00e9ines humaines qui sont cibl\u00e9es par la SidJ.<\/p>\n\n\n\n

Toxine au travail<\/strong><\/h3>\n\n\n\n

Le groupe a montr\u00e9 que la SidJ poss\u00e8de une activit\u00e9 de glutamylation des prot\u00e9ines : elle attache le glutamate d’acide amin\u00e9 \u00e0 une prot\u00e9ine cible comme modification post-traductionnelle. “Ce type d’activit\u00e9 est une premi\u00e8re pour les prot\u00e9ines bact\u00e9riennes “, d\u00e9clare Ivan Dikic, directeur de l’Institut de biochimie II de l’Universit\u00e9 Goethe. La SidJ glutamylate de nombreuses prot\u00e9ines humaines impliqu\u00e9es dans la lutte contre les infections microbiennes et l’immunit\u00e9 inn\u00e9e. Pour ce faire, la SidJ interagit avec la prot\u00e9ine humaine Calmodulin – une prot\u00e9ine messag\u00e8re interm\u00e9diaire multifonctionnelle de liaison au calcium hautement conserv\u00e9e. “La Legionella<\/em> a intelligemment \u00e9volu\u00e9 pour utiliser la Calmoduline pour d\u00e9clencher l’activit\u00e9 de SidJ et, par cons\u00e9quent, emp\u00eacher l’activation de SidJ avant que l’infection dans le corps humain n’ait lieu”, dit Dikic.<\/p>\n\n\n\n

La structure de microscopie \u00e9lectronique cryog\u00e9nique de SidJ interagissant avec la calmoduline humaine a \u00e9galement r\u00e9v\u00e9l\u00e9 que la toxine a un pli de domaine kinase. “C’est une d\u00e9couverte \u00e0 la fois int\u00e9ressante et importante, car le pli kinase est droguable “, dit Michael Adams, un \u00e9tudiant en doctorat du groupe Bhogaraju.<\/p>\n\n\n\n

D\u00e9but d’un long chemin vers l’utilisation th\u00e9rapeutique<\/strong><\/h3>\n\n\n\n

Les r\u00e9sultats de l’\u00e9tude vont amorcer de nombreuses \u00e9tudes \u00e0 l’avenir, diss\u00e9quant davantage le m\u00e9canisme de la glutamylation \u00e0 m\u00e9diation de SidJ. Fait important, puisque les chercheurs ont d\u00e9couvert que la SidJ a un pli kinase, cette d\u00e9couverte amorcera la recherche d’une mol\u00e9cule m\u00e9dicamenteuse ayant des effets th\u00e9rapeutiques potentiels.<\/p>\n\n\n\n

“Bien que nos travaux n’aient pas d’application pharmaceutique directe, nos r\u00e9sultats sur les caract\u00e9ristiques structurelles et fonctionnelles d’une des toxines les plus importantes de Legionella<\/em>, conduiront \u00e0 des \u00e9tudes futures visant \u00e0 cibler cette prot\u00e9ine \u00e0 des fins th\u00e9rapeutiques”, d\u00e9clare Sagar Bhogaraju.<\/p>\n","protected":false},"excerpt":{"rendered":"

SidJ enforces a unique modification on human proteins and helps legionella grow.<\/p>\n","protected":false},"author":71,"featured_media":16765,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[2,17591],"tags":[789,37,3684,893,1748,765],"embl_taxonomy":[],"class_list":["post-16763","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-science","category-science-technology","tag-bhogaraju","tag-grenoble","tag-infection-biology","tag-legionella","tag-press-release","tag-toxin"],"acf":{"article_intro":"","related_links":[{"link_description":"Bhogaraju Group (EMBL Grenoble)","link_url":"https:\/\/www.embl.fr\/research\/unit\/bhogaraju\/index.html"},{"link_description":"Molecular Signaling Group (Goethe Univerity Frankfurt)","link_url":"https:\/\/www.biochem2.com\/index.php\/9-ibcii\/msg\/93-msg-frontpage"}],"article_sources":[{"source_description":"

Sagar Bhogaraju, Florian Bonn, Rukmini Mukherjee, Michael Adams, Moritz M. Pfleiderer, Wojciech P. Galej, Vigor Matkovic, Sissy Kalayil, Donghyuk Shin, Ivan Dikic: Inhibition of SidE ubiquitin ligases through SidJ\/Calmodulin catalyzed glutamylation. 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