Thesis (M.S.)--Michigan State University. Dept. of Fisheries and Wildlife, 2008. / Title from PDF t.p. (viewed on July 29, 2009) Includes bibliographical references (p. 64-73). Also issued in print.

Thesis (M.D.)--Universitet Københavns, 2006. / Includes bibliographical references.

Approximately 2-4 billion cases of infectious diarrhoea occur every year, with the highest numbers recorded in sub-Saharan Africa. It remains the most common public health issue among children in developing nations. The purpose of this research was to unfold the prevalence of diarrhoeagenic E. coli and Campylobacter pathotypes as well as elucidate their antibiogram characteristics in diarrhoeal stool samples collected in some medical facilities in Eastern Cape Province, South Africa. Two hundred stool samples were collected from both inpatients and outpatients from male and females of all age groups attending selected medical facilities in the study area. Isolation and characterization of both organisms were done using culture based and molecular methods. Antibiotic susceptibility patterns of identified isolates were determined against a panel of 12 antimicrobial agents. One hundred and twenty presumptive E. coli isolates and 42 presumptive isolates of Campylobacter spp. Were isolated. Eighty-two percent (82 percent) of the presumptive E. coli isolates were confirmed as E. coli while 46.3 percent belonged to Campylobacter spp. Pathotyping of the diarrhoeagenic E. coli isolates by Polymerase chain reaction (PCR) showed the following prevalences: DAEC 43 (32 percent), EHEC 18 (17 percent), EIEC 11 (10 percent) and EPEC 18 (17 percent). EAEC and ETEC were not detected, while for Campylobacter spp. 37 (88 percent) were C. jejuni, and C. coli was not detected. A total of 12 (32.4 percent) of the confirmed Campylobacter jejuni isolates were found to possess the fliM gene, 9 (24.3 percent) possessed the flhA gene and only 6 (16.2 percent) harboured the gene flgE2. None were positive for the flaA, flab and flhB genes.The antibiotic resistance patterns observed among the E. coli isolates were high against ampicillin (98.1 percent), chloramphenicol (94.3 percent) and tetracycline (90.6 percent). For Campylobacter spp., resistance observed were: chloramphenicol (91.6 percent), tetracycline (25.2 percent), erythromycin (49.6 percent) and gentamycin (56.4 percent). A lesser resistance against imipenem (35.9 percent) and quinolone (ciprofloxacin) (45.5 percent) were exhibited by the E.coli isolates. 10.8 percent and 20.3 percent of the Campylobacter isolates were resistant to imipenem and ciprofloxacin respectively. The presence of chloramphenicol (CatA1) and tetracycline (tetA) resistance genes were detected in 94 percent and 89 percent of E. coli isolates respectively while 98 percent of Campylobacter spp. Harboured the catA1 resistance gene. It could be deduced from this study that E. coli and Campylobacter spp. are predomiant enteric pathogens as the etiologic agents of diarrhoea in the study community, and that their antimicrobial resistance is high in the study location. The need to develop strategies to prevent infection and control resistant organisms is evident.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Campylobacter sp. à uma importante causa de enterite de origem alimentar com alta incidÃncia na populaÃÃo infantil de paÃses em desenvolvimento. No entanto, o diagnÃstico especÃfico de sua etiologia segue como um desafio, visto que mÃtodos moleculares e imunoenzimÃticos tÃm se mostrado mais sensÃveis. Postulamos que o conhecimento de sua virulÃncia e o diagnÃstico especÃfico possam ajudar na identificaÃÃo e potencial controle da campilobacteriose na infÃncia. Foi determinada a etiologia de diarreia por Campylobacter sp., em um estudo transversal sobre diarreia em crianÃas de 0-36 meses residentes da Ãrea urbana de Fortaleza, CearÃ, Brasil, que necessitaram de atendimento mÃdico de urgÃncia por causa de doenÃa diarreica. ApÃs a aprovaÃÃo Ãtica do estudo, um questionÃrio foi aplicado para qualificar as condiÃÃes clÃnicas apresentadas por cada crianÃa no momento da admissÃo. O DNA foi extraÃdo diretamente de amostras fecais coletadas de 226 crianÃas. Para a detecÃÃo do agente etiolÃgico, utilizamos diagnÃstico molecular (PCR e RT-PCR) e diagnÃstico imunoenzimÃtico (ELISA), alÃm da detecÃÃo de genes associados à virulÃncia de C. jejuni (PCR). Campylobacter sp. foi encontrado em 8,9% (20/225) das amostras, por diagnÃstico microbiolÃgico convencional. C. jejuni e C. coli foram detectados em 19,5% (44/226) e 1,3% (3/226) das amostras diarreicas, respectivamente, por PCR. Os diagnÃsticos por RT-PCR e ELISA alcanÃaram 26,7% (60/225) e 37,9% (58/153), respectivamente. Quando considerada a combinaÃÃo de diagnÃsticos (positividade no diagnÃstico microbiolÃgico ou no imunoenzimÃtico e ao menos em um dos testes moleculares) a prevalÃncia encontrada foi de 16,4% (37/226). A concordÃncia entre os testes para diagnÃstico utilizados foi de moderada a regular, de acordo com o Ãndice de Kappa. Genes associados à virulÃncia foram detectados nas seguintes proporÃÃes de amostras positivas para C. jejuni: flaA, 79,5% (35/44); racR, 97,7% (43/44) e dnaJ, 88,6% (39/44) â relacionados à adesÃo bacteriana e colonizaÃÃo; ciaB, 97,7% (43/44); pldA, 45,4% (20/44) e pVir 0% (0/44) â relacionados à invasÃo; e cdtABC em 95,4% (42/44) das amostras, operon relacionado à produÃÃo da toxina citoletal distensora (CDT). Sinais e sintomas especÃficos, tais como sangue nas fezes, vÃmito, febre e/ou dor abdominal, apesar de bastante frequentes, nÃo foram associados com a detecÃÃo de C. jejuni. O perfil de distribuiÃÃo dos genes de virulÃncia de C. jejuni nÃo apresentou correlaÃÃo com a apresentaÃÃo clÃnica da doenÃa, mesmo quando tal perfil foi categorizado de acordo com a funÃÃo das proteÃnas codificadas pelos genes, o que nos leva a crer que outros fatores, talvez relacionados à susceptibilidade do hospedeiro, possam ser mais importantes do que a variabilidade genÃtica do micro-organismo. ConcluÃmos que Campylobacter sp. foi detectado em percentual relevante da populaÃÃo estudada, principalmente quando os mÃtodos diagnÃsticos foram utilizados de forma combinada. Em geral, os genes de virulÃncia foram detectados em uma alta proporÃÃo das amostras positivas para C. jejuni, embora os genes relacionados à invasÃo tenham sido menos frequentemente encontrados. Corroboramos dados de outros grupos sobre a necessidade de revisÃo do diagnÃstico para Campylobacter sp. em prol da inclusÃo de metodologias mais sensÃveis e espÃcie-especÃficas, alÃm da busca por marcadores para inflamaÃÃo intestinal e fatores preditivos de cultura negativa. / Campylobacter sp. is an important cause of food-borne gastroenteritis with high incidence in children living in developing countries. However, the specific diagnosis of its etiology remains as a challenge, since conventional diagnosis by culture is now challenged by molecular and immunoenzymatic methods, which have greater sensitivity. We postulate that the knowledge of its virulence and specific diagnosis may assist in identifying and potentially controling campylobacteriosis in childhood. We determined the etiology of Campylobacter sp. associated diarrhea, in a cross-sectional study of diarrhea in children aged 0-36 months from the urban area of Fortaleza, CearÃ, Brazil, who required emergency medical care because of diarrheal disease. After ethical approval of the study, a questionnaire was applied to describe the clinical conditions presented by each child at the time of admission. DNA was extracted directly from fecal samples collected from 226 children. For the determination of the etiologic agent we used molecular diagnostics (PCR and RT-PCR) and diagnostic immunoassay (ELISA), besides the detection of virulence associated genes of C. jejuni (PCR). Campylobacter sp. was found in 8.9% (20/225) of the samples by conventional microbiological diagnosis. C. jejuni and C. coli were detected in 19.5% (44/226) and 1.3% (3/226) of the diarrheic samples, respectively. The diagnostic RT-PCR and ELISA reached 26.7% (60/225) and 37.9% (58/153) of positivity, respectively. When considering the combination of diagnostic (positive in microbiological diagnosis or immunoassay and at least one of the molecular tests) the prevalence was 16.4% (37/226). The agreement between the tests used for diagnosis was moderate to regular, according to Kappa index. The presence of C. jejuniÂs virulence-associated genes that encode proteins related to the pathogenesis of micro-organism were detected in the following proportions of C. jejuni-positive DNA samples: flaA, 79.5% (35/44); racR, 97.7% (43/44) and dnaJ, 88.6% (39/44) â related to bacterial adhesion and colonization; ciaB, 97.7 % (43/44); pldA, 45.4% (20/44) and pVir 0% (0/44) â related to invasion, and cdtABC in 95.4% (42/44) of samples related to citoletal distending toxin (CDT). Specific signs and symptoms such as blood in the stool, vomiting, fever and/or abdominal pain, although quite frequent, were not associated with the detection of C. jejuni. The distribution profile of C. jejuniÂs virulence genes was not correlated with the clinical presentation of the disease, even when this profile was categorized according to the function of the proteins encoded by the genes, which leads us to believe that other factors, perhaps related to host susceptibility, may be more important than genetic variability of the microorganism. We conclude that Campylobacter sp. was detected in a significant percentage of the children 0-36 months with diarrhea, especially when the diagnostic methods were used in combination. In general, the virulence genes were detected in a high proportion of C. jejuni-positive samples, although the invasion-related genes have been found less frequently. Our data corroborates findings from other groups on the need to revise the diagnostic for Campylobacter sp. towards the inclusion of more sensitive and species-specific methods, as well as search for extra markers for intestinal inflammation and predictors of negative culture.

Campylobacter jejuni is an avian commensal bacterium and causes gastrointestinal diarrhea in humans called campylobacteriosis. Campylobacteriosis is acquired by consumption of undercooked poultry contamined with C. jejuni. Poultry can become colonized from contaminated drinking water. The chicken flock and drinking water of 4 poultry farms in Ontario were sampled and the prevalence of C. jejuni in these flocks was determined to be 16.7% over a 1 year sampling period. We determined that contamined- water was a significant risk factor for Campylobacter-positive flocks from flaA typing, PFGE analysis, and genomotyping several isolated strains. Free living amoebae, such as Acanthamoeba species, live in the drinking water of poultry farms. It is hypothesized that Acanthamoeba in the drinking water of poultry farms can take up and act as environmental reservoirs of C. jejuni. Acanthamoeba species were isolated from the drinking water. Acanthamoeba strains were found to act as a vehicle for protection, persistence and growth of C. jejuni isolated from the farm water. The transcriptome of both C. jejuni and A. castellanii during the initial stages of C. jejuni internalization were described by RNA-seq. C. jejuni oxidative defence genes (such as katA, sodB, fdxA) and some other unknown genes (Cj0170, Cj1325, Cj1725) were found to be essential in the interaction with A. castellanii. Our findings suggest that Acanthamoebae act as a C. jejuni reservoir and could be a contributing source of C. jejuni in the environment. Through transcriptomics studies, we have begun to uncover some genetic clues involved in this interaction.

Campylobacter jejuni is a leading cause of food poisoning and according to the World Health Organisation accounts for majority of the 4.5 billion cases of global food poisoning each year. Genome sequencing by Parkhill et al. (2000) identified a gene, cj1411c, which is thought to encode a lone cytochrome P450, CYP172A1. In this thesis the role of CYP172A1 was studied using in vivo and in vitro techniques. The genomic location of cj1411c is adjacent to the capsular biosynthetic genes. The capsular and P450 genes are conserved in some species of Campylobacter and Helicobacter, as well as in Comamonas testosteroni. Importantly, this work has demonstrated that the P450 gene is expressed in two well characterised laboratory C. jejuni strains, 11168H and 81-176. Protein production was disrupted using insertional knockout mutagenesis, which allowed for investigations into the role of the enzyme in the host. Alterations to the observed autoagglutination rate and growth characteristics indicated that CYP172A1 has a role in modifying the bacterial surface. The insertional knockout mutant also resulted in cells which were more susceptible to detergent-like compounds (e.g. polymyxin B and sodium deoxycholate). In a previous report, it was suggested that the loss of the P450 function resulted in bacteria which were “shorter and fatter”, compared to wild type cells, but this thesis could find no evidence of such a phenomenon. CYP172A1 was successfully purified using recombinant expression in E. coli to enable biochemical and biophysical characterisation in vitro. CYP172A1 contains a typical P450 cysteine thiolate coordination to the heme iron, and exists in a low spin ferric heme state under neutral buffer conditions. The P450 was found to self aggregate, and despite rigorous investigations the cause of this aggregation was not fully established. Despite this issue, CYP172A1 was shown to bind to a wide range of P450 inhibitor-type compounds, with econazole displaying the tightest binding affinity (Kd = 100 nM). Identification of substrate-like compounds was achieved using high throughput compound screening, and a number of organic compounds were identified and shown to bind CYP172A1, inducing heme iron absorbance changes typical of either P450 inhibitors or substrates. Optical titrations for these molecules indicated that their CYP172A1 Kd values were in the low micromolar range. The catalytic capability of CYP172A1 was successfully demonstrated by providing the P450 with non native redox partners to oxidise one of such substrate-like compound (213071), resulting in the sulfoxidation of this compound.

Campylobacteriosis is according to European Food Safety Authority one of the most advanced worldwide foodborne disease causing gastroenteritis. The Czech republic ranks at number one in the highest incidence of campylobacteriosis. This thesis deals with the determination of thermotolerant Campylobacter spp. in samples of poultry and pork liver, in chicken and raw milk. From total number of 50 samples were 31 samples positive for Campylobacter spp. For poultry liver and chicken at the back of thighs were 92 % of positive samples. For pig liver was lower incidence, only 46 %. For fresh raw milk was the lowest incidence of just 17 %. Higher incidence in the Czech republic reflects good way of searching their subsequent reporting infections.

The objective of this dissertation was to (1) determine which grow-out and processing sampling points best predicts and causes Campylobacter later in production (2) identify risk factors within the hatchery that influenced Campylobacter prevalence later in production (3) identify biosecurity risk factors that were associated with Campylobacter presence during production and processing (4) identify farm and production characteristics that were associated with Campylobacter presence later in production, and (5) to estimate the proportion of variance and the intraclass correlation coefficients within the hierarchical levels (complex, farm, bird) of the data. The best predictors of post-chill Campylobacter carcass status were the exterior whole carcass sample in the grow-out environment and the crop upon arrival at the processing plant. The best post-chill causal model contained the grow-out whole carcass. Variables associated with the increased odds of a Campylobacter positive sample included controlling the humidity in the hatchery chick room, 2-4 people handling the chicks at the hatchery, washing the setter twice yearly, 2 or more breeder farms providing eggs for the sampled flock, using low water pressure when washing the hatch trays, having more walk-in doors on the boiler house, the farmer removing the litter from the farm, concrete at most used door of the broiler house, the number of workers that work with the birds during grow-out, having more houses on the farm, standing water on the farm day 1, wood interior walls, a vegetation surface next the house footing, and 6 or less flocks on the litter. Protective factors included the use of footbaths and dedicated shoes, greater frequency of entering the house during brooding, disinfectant added to the drinker lines, having concrete outside the most used door, the cleanliness of the workroom, and harvesting birds at 56-63 days of age. The highest percent of variance occurred at the farm level meaning intervention efforts should focus on factors at the broiler farm level i.e. factors that are different among farms within a broiler complex.

Campylobacter is the leading cause of bacterial gastroenteritis in the United States. The objective of this study was to investigate the prevalence of Campylobacter in newly constructed broiler houses and compare three microaerophilic gas delivery methods used to culture Campylobacter in the laboratory. Of 2,300 litter, 900 fecal, and 45 water samples, only 5, 6 and 1 of the samples, respectively, were confirmed positive. style='mso-spacerun:yes'> Results indicated litter moisture content was different across day, location and house. An interaction was detected for litter pH between day, location and flock. Temperatures averaged 26.8°C inside and 27.6°C outside. style='mso-spacerun:yes'> No difference in colony counts were detected among the gas delivery methods. In conclusion, the newly constructed houses showed no significant prevalence of Campylobacter. style='mso-spacerun:yes'> High litter pH, low temperatures, and other onarm management strategies may have suppressed Campylobacter’s ability to colonize the litter. When selecting a gas delivery method price and space should be considered.

More than 150 different RNA modifications have been detected in all kingdoms of life and 60 are known to decorate bacterial RNA. Among them, pseudouridine is universally conserved and one of the most abundant modifications present in bacterial stable RNAs such as tRNAs and rRNAs. In bacteria, the nucleotide is posttranscriptionally generated by dedicated enzymes called pseudouridine synthases (PUSs). With the advent of sophisticated deep-sequencing technologies, this modification has been identified in different types of RNA classes (tRNAs, rRNAs, mRNAs, snRNAs, and lncRNAs) in diverse eukaryotic organisms. However, these techniques have never been applied to bacteria, generating a knowledge gap about the location of the modified nucleotide in prokaryotic RNAs. Mutations or deletions of specific eukaryotic PUS enzymes are linked to human diseases and therefore their absence is deleterious for the correct function of the cell. However, deletion of tRNA or rRNA PUS enzymes in the bacterial model organism E. coli have not revealed any such drastic phenotypes, suggesting a different role and function of the modification itself and of the enzymes in different kingdoms of life. Since the roles of tRNA PUS enzymes in bacteria is still poorly understood, a functional characterization of these proteins is pursued in the Epsilonproteobacteria Campylobacter jejuni and Helicobacter pylori. While C. jejuni is the leading cause of bacterial foodborne gastroenteritis in humans, infection with H. pylori is associated with the development of gastric cancer. In particular, phenotypes were explored for the tRNA PUS enzymes TruA, TruB, and TruD in C. jejuni as well as TruA and TruD in H. pylori. Upon deletion of truD, a severe growth defect is observed for C. jejuni but not for H. pylori, highlighting a potential difference in function of the enzyme in the two related bacterial pathogens. Moreover, a genome-wide approach called Pseudo-seq is established and applied for RNA of these two pathogens, which allows, for the first time, the global identification of pseudouridine modifications at single-nucleotide resolution in the bacterial transcriptome. Applying Pseudo-seq in RNAs of wildtype and diverse PUS enzyme deletion mutants enabled the identification of the distinct RNA substrates of tRNA PUS enyzmes in C. jejuni and H. pylori. Hereby, the tRNA-Glu was determined to be the major tRNA substrate of TruD in C. jejuni. Interestingly, the tRNA-Glu is expressed as a single copy in the C. jejuni genome. To link the growth defect observed for a C. jejuni ∆truD mutant strain to the pseudouridine modification of the tRNA-Glu, a catalytically inactive TruD complementation was generated. This strain is unable to restore the tRNA-Glu modification but surprisingly, was able to complement the growth defect. The same observation was made for a cross-complementation with a copy of H. pylori TruD. This indicates that there is a potential additional function of the TruD PUS enzyme in C. jejuni that is independent of the pseudouridine modification. Using a combination of deep-sequencing technologies (RIP-seq, RNA-seq, Ribo-seq, and CLIP-seq), the dual function of TruD is investigated. Overall, this study provides the first in-depth investigation into pseudouridylation of bacteria in general and the bacterial pathogens C. jejuni and H. pylori in particular. The work presented in this thesis reveals not only a global map of pseudouridine in tRNAs and rRNAs of the two bacteria but it also explores the function of the responsible tRNA PUS enzymes. In addition, this study provides evidence for a dual function of the C. jejuni PUS enzyme TruD that goes beyond its RNA modifying function. Future research could focus on unravelling the function of TruD and its potential interaction partners and thus reveal new mechanisms of regulation of a protein previously only described as an RNA modification enzyme. / Mehr als 150 verschiedene RNA-Modifikationen sind bislang in den unterschiedlichsten Organismen nachgewiesen worden, wovon 60 dieser Modifikationen in bakterieller RNA vorkommen. In Bakterien ist Pseudouridin eine der häufigsten Modifikationen, die in stabilen RNAs wie tRNAs und rRNAs zu finden sind. Hierbei wird das modifizierte Nukleotid auf posttranskriptioneller Ebene von speziellen Enzymen, den sogenannten Pseudouridin-Synthasen (PUS), generiert. Die Entwicklung und der Einsatz fortschrittlicher Deep-Sequencing Technologien ermöglichte es, Pseudouridin in unterschiedlichen RNA Klassen (tRNAs, rRNAs, mRNAs, snRNAs und lncRNAs) in verschiedenen eukaryotischen Organismen zu identifizieren. Diese Verfahren wurden jedoch noch nie auf Bakterien angewandt. Mutationen oder Deletionen spezifischer PUS Enzyme wurden im Menschen bereits mit der Entstehung von Krankheiten in Verbindung gebracht. Diese Enzyme sind daher für die korrekte Funktionsweise einer eukaryotischen Zelle unabdinglich. Nichtsdestotrotz führte die Deletion von tRNA oder rRNA PUS Enzymen im bakteriellen Modellorganismus Escherichia coli zu keinen solch drastischen Phänotypen. Dies wiederum deutet auf eine unterschiedliche Rolle und Funktion der Modifikation und der verantwortlichen Enzyme in verschiedenen Organismen hin. In der vorliegenden Arbeit werden tRNA PUS Enzyme der Epsilonproteobakterien Campylobacter jejuni und Helicobacter pylori funktionell charakterisiert. Der Lebensmittelkeim C. jejuni ist derzeit die häufigste Ursache für bakteriell verursachte Gastroenteritis im Menschen. Dahingegen wird eine H. pylori Infektion mit der Entwicklung von Magenkrebs in Verbindung gebracht. Insbesondere wurden die Funktionen der tRNA PUS Enzyme TruA, TruB und TruD in C. jejuni sowie TruA und TruD in H. pylori untersucht. Während die Deletion von TruD keine phänotypischen Auswirkungen in H. pylori hat, führt diese in C. jejuni zu einem Wachstumsdefekt. Dies weist auf eine möglicherweise unterschiedliche Funktion des Enzyms in den beiden verwandten bakteriellen Krankheitserregern hin. Zusätzlich beschreibt diese Arbeit die Etablierung und Anwendung von Pseudo-seq in C. jejuni und H. pylori, einem genomweiten Ansatz mittels dessen zum ersten Mal die globale Identifizierung von Pseudouridin Modifikationen auf Einzel-Nukleotid-Ebene im bakteriellen Transkriptom ermöglicht wird. Durch Pseudo-seq Analysen von wildtypischer RNA und RNA isoliert aus unterschiedlichen PUS Enzym Deletionen, konnten die RNA Substrate dieser Enzyme in C. jejuni und H. pylori ermittelt werden. Für TruD stellte sich dabei die tRNA-Glu als Hauptsubstrat heraus. Interessanterweise ist diese im Genom von C. jejuni nur als einzelne Kopie vorhanden. Da eine TruD Deletionsmutante in C. jejuni einen Wachstumsdefekt aufweist, wurde dieser Phänotyp in Zusammenhang mit dem Auftreten der Pseudouridin Modifikation an der tRNA-Glu untersucht. Zu diesem Zweck wurde ein TruD Komplementationsstamm generiert, der jedoch katalytisch inaktiv ist und somit nicht in der Lage ist die Modifikation der tRNA-Glu wiederherzustellen. Überraschenderweise komplementierte dieser Stamm dennoch den Wachstumsdefekt. Eine ähnliche Beobachtung wurde bei einer Kreuzkomplementation mit einer Kopie von H. pylori TruD gemacht. Dies deutet darauf hin, dass das TruD PUS Enzym in C. jejuni möglicherweise eine zusätzliche Funktion unabhängig von der Pseudouridin Modifikation hat. Diese potentiell duale Funktion von TruD wird in dieser Arbeit durch die Anwendung einer Kombination von Deep-Sequencing Technologien (RIP-seq, RNA-seq, Ribo-seq und CLIP-seq) untersucht. Insgesamt stellt diese Studie die erste eingehende Untersuchung von Pseudouridin Modifikationen in Bakterien im Allgemeinen, und in den Krankheitserregern C. jejuni und H. pylori im Speziellen, dar. Die in dieser Arbeit vorgelegten Ergebnisse beschreiben nicht nur eine globale Kartierung von Pseudouridin Modifikationen in bakteriellen tRNAs und rRNAs sondern erforschen auch die Funktionen der für die Modifikation verantwortlichen tRNA PUS Enzyme. Darüber hinaus liefert diese Arbeit Hinweise auf eine duale Funktion des C. jejuni PUS Enzyms TruD, die über die Funktion als RNA-modifizierendes Enzym hinausgeht. Zukünftige Untersuchungen könnten sich dementsprechend darauf konzentrieren, die Funktion von TruD und seinen potenziellen Interaktionspartnern zu entschlüsseln. Dies könnte neue Erkenntnisse über Mechanismen der Regulierung eines Enzyms/Proteins liefern, das bislang nur als RNA modifizierendes Enzym beschrieben war.