Human histidyl-tRNA synthetase (HARS) is an aminoacyl-tRNA synthetase (AARS) that catalyzes the attachment of the amino acid histidine to histidyl-tRNA (tRNAHis) in a two-step reaction that is essential for protein translation. Currently, two human diseases, Usher Syndrome IIIB (USH3B) and an inherited peripheral neuropathy, Charcot Marie Tooth Syndrome (CMT), have been linked genetically to single point mutations in the HARS gene. The recessive HARS USH3B mutation encodes an Y454S substitution localized at the interface between the anticodon-binding domain and the catalytic domain of the opposing subunit. Patients with Usher Syndrome IIIB lose their sight and hearing during their second decade of life, and clinicians have observed that the onset of deafness and blindness may be episodic and correlate with febrile illness. Furthermore, some young USH3B patients present with a fatal form of acute respiratory distress. In addition to the single HARS mutation linked to Usher Syndrome, eight other mutations in the HARS gene are associated with CMT, an inherited peripheral neuropathy. Peripheral neuropathies are associated with progressive and length-dependent damage of the motor and sensory neurons that transmit information to the spinal cord. The age of onset and phenotypic severity of CMT linked to HARS is highly variable. When expressed in a yeast model system, the HARS variants are dominantly lethal, and confer defects in axonal guidance and locomotor deficiencies when expressed in C.elegans. Here, the biochemical characterization of the HARS USH3B and three peripheral neuropathy variants are described. The approaches included enzyme kinetic analysis with purified HARS enzymes to monitor catalytic deficiencies, differential scanning fluorimetry (DSF) to evaluate structural instability, and cellular models to detect physiological effects of axonal outgrowth by CMT variants. The results suggest that Usher Syndrome IIIB is unlikely to be a consequence of a simple loss of aminoacylation function, while HARS-linked peripheral neuropathy variants all share common catalytic defects in aminoacylation. The HARS system represents a notable example in which two different complex human diseases arise from distinct mutations in the same parent gene. By understanding the biochemical basis of these inherited mutations and their link to Usher Syndrome and CMT, it may be possible to develop mechanism-based therapies to improve the quality of life of patients afflicted with them.

Les photorécepteurs sont des neurones très spécifiques dédiés à la phototransduction et reposant sur une machinerie cellulaire très complexe. La dépolarisation permanente dans le noir des photorécepteurs déclenche une transmission synaptique constante et extrêmement spécifique qui requièrent une quantité d'énergie considérable. Les photorécepteurs peuvent dégénérer lorsque la phototransduction ou l'apport énergétique sont altérés. Le syndrome d'Usher conduit à une surdité et une cécité. La recherche du rôle des protéines usher dans les photorécepteurs a été freinée par l'absence de phénotype rétinien dans les modèles. De la même façon, la compréhension des mécanismes moléculaires conduisant à l'atteinte des cônes dans la rétinopathie diabétique a été entravée par l'absence de symptômes vasculaires et neuronaux dans les modèles. Durant ma thèse, j'ai caractérisé deux modèles animaux des syndromes Usher et HANAC. Des atteintes neuronales ont été démontrées par électrorétinogramme et par l'observation de changements morphologiques des cellules. Dans les modèles Usher, j'ai également montré une neuroprotection des photorécepteurs par plusieurs stratégies. Dans le modèle HANAC, les atteintes neuronales étaient associées à une tortuosité vasculaire anormale une augmentation de la perméabilité vasculaire et l'expression accrue de VEGF. Les évaluations phénotypiques de ces trois modèles fournissent un nouvel aperçu de la physiopathologie des dégénérescences des cônes dans le syndrome d'Usher et dans les maladies vasculaires complexes. Ce travail ouvre surtout la voie au développement et à l'évaluation de nouvelles stratégies thérapeutiques pour ces maladies menant à la cécité. / Photoreceptors are very specific neurons dedicated to phototransduction, which relies on very complex machinery. The maintained depolarization in darkness triggers a constant and thus very specific type of synaptic transmission. These require high energy need. As a consequence, photoreceptors can degenerate in various hereditary retinal diseases when phototransduction or energy consumption are altered. The Usher syndrome is such a hereditary disease leading to both deafness and blindness. If Usher proteins are involved in the mechanotransduction in hair cells, investigating their role in photoreceptors has been hamperedby the lack of a retinal phenotype in murine models. Similarly, understanding themolecular mechanisms of cone dysfunction in diabetic retinopathy has beenhampered by the lack of vascular and neuronal symptoms and neuronal models. During my PhD, I have developed animal models of Usher and HANAC syndromes both leading to cone photoreceptor dysfunction and damage. Cone dysfunction was demonstrated by electroretinogram recording and by morphological changes, retinal gliosis and microglial activation. In the Usher models, I also demonstrated photoreceptor neuroprotection by different strategies. In the HANAC model, neuronal dysfunction was associated as in diabetic retinopathy to blood vessel tortuosity, blood vessel permeability and incresead VEGF expression levels. These phenotypic evaluations of mouse models provide new insights into the physiopathology of cone photoreceptor degeneration in Usher syndrome and in complex vascular diseases. It also open the way for the development and assessment of new therapeutic strategies for these diseases leading to blindness.

No description available.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20789号 / 医博第4289号 / 新制||医||1025(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 山田 亮, 教授 大森 孝一, 教授 高田 穣 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

The very large G-protein-coupled receptor 1 (VLGR1/ADGRV1) is the largest member of the adhesion G-protein-coupled receptor (ADGR) family. Mutations in VLGR1/ADGRV1 cause human Usher syndrome (USH), a form of hereditary deaf-blindness, and have been additionally linked to epilepsy. In the absence of tangible knowledge of the molecular function and signaling of VLGR1, the pathomechanisms underlying the development of these diseases are still unknown. Our study aimed to identify novel, previously unknown protein networks associated with VLGR1 in order to describe new functional cellular modules of this receptor. Using affinity proteomics, we have identified numerous new potential binding partners and ligands of VLGR1. Tandem affinity purification hits were functionally grouped based on their Gene Ontology terms and associated with functional cellular modules indicative of functions of VLGR1 in transcriptional regulation, splicing, cell cycle regulation, ciliogenesis, cell adhesion, neuronal development, and retinal maintenance. In addition, we validated the identified protein interactions and pathways in vitro and in situ. Our data provided new insights into possible functions of VLGR1, related to the development of USH and epilepsy, and also suggest a possible role in the development of other neuronal diseases such as Alzheimer’s disease.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-02T13:12:16Z No. of bitstreams: 2 Dissertação - Ricardo Campos Cruvinel Filho - 2014.pdf: 8513057 bytes, checksum: e42a2106d7e7abda6b1ee9d65068229d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-02T13:28:23Z (GMT) No. of bitstreams: 2 Dissertação - Ricardo Campos Cruvinel Filho - 2014.pdf: 8513057 bytes, checksum: e42a2106d7e7abda6b1ee9d65068229d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-08-02T13:28:23Z (GMT). No. of bitstreams: 2 Dissertação - Ricardo Campos Cruvinel Filho - 2014.pdf: 8513057 bytes, checksum: e42a2106d7e7abda6b1ee9d65068229d (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-12-16 / Cross-sectional study conducted at the Center of Reference in Ophthalmology UFG in conjunction with Oregon Health and Science University and the Brazilian Center for Eye Surgery (CBCO). To evaluate the genotype of patients with Usher syndrome of Reference Center for Ophthalmology, Federal University of Goias (UFG-CEROF). Patients clinically diagnosed with SU underwent complete ophthalmic examination, Goldmann manual kinetic perimetry, audiometry and subsequent collection of peripheral blood chromosomal microarray for sequencing. We examined 19 patients with clinical suspicion of SU with a mean age at first visit was 42.5 years (± 12.2) and a slight predominance of males (52.63%). The most prevalent subtype in clinical diagnosis of type I disease (68.4%). The visual acuity measured on the day of the exam for eye examination was 20/92 on the Snellen chart. Examinations audiometry showed hearing loss in all patients ranging from moderate in 12.5% of patients, deep (56.25%) and severe (31.25%). In 36.8% of patients analyzed, we found at least two mutations in the same gene, and of these, 21% were heterozygous mutations, and 15.8% homozygous. The homozygous mutations, which were of the type no sense, occurred in the gene CLRN1 whose patients had a previous diagnosis of USH 2. Met 26.31% of the sample analyzed in heterozygous. Of these, two patients showed mutations in the MYO7A gene (40%), both with clinical suspicion of USH 1. For the proposed methodology, we found no disease-causing mutations in 79% of the sample analyzed. Following the proposed methodology, the authors were able to determine the mutation in seven patients of nineteen patients inclued in this study. Of these, three patients were diagnosed with homozygous mutations in gene CLRN1, and had previous clinical diagnosis of type 2. Two patients had heterozygous mutations in gene MYO7A, both with previous clinical diagnosis of type 1. / Estudo transversal, desenvolvido no Centro de Referência em Oftalmologia da UFG em conjunto com a Oregon Health and Science University e Centro Brasileiro de Cirurgia de Olhos (CBCO), que teve como objetivo avaliar o genótipo de pacientes com síndrome de Usher do Centro de Referência em Oftalmologia da Universidade Federal de Goias (CEROF-UFG). Pacientes clinicamente diagnosticados com SU foram submetidos a exame oftalmológico completo, perimetria cinética manual de Goldmann, audiometria e posterior coleta de sangue periférico para sequenciamento cromossômico por microarray. Foram examinados 19 pacientes com diagnostico clínico de SU com média de idade na primeira consulta de 42,5 anos (± 12,2) e pequena predominância do sexo masculino (52,63%). O subtipo mais prevalente no diagnóstico clínico foi do tipo I da doença (68,4%). A acuidade visual média medida no dia do exame por olho examinado foi de 20/92 na escala de Snellen. Os exames audiométricos mostraram perda de audição em todos pacientes variando de moderada em 12,5% dos pacientes, profunda (56,25%) e severa (31,25%). Em 36,8% dos pacientes analisados, encontraram-se ao menos duas mutações em um mesmo gene, sendo que destes, 21% eram mutações heterozigotas e, 15,8% homozigotas. As mutações homozigotas, as quais eram do tipo sem senso, ocorreram no gene CLRN1, cujos pacientes tinham o diagnóstico clínico prévio de USH 2. Encontrou-se 26,31% da amostra analisada em heterozigose. Desses, dois pacientes mostraram mutações para o gene MYO7A (40%), ambos com suspeita clínica de USH 1. Pela metodologia proposta, não foram encontradas mutações causadoras de doença em 79% da amostra analisada. Dos 19 pacientes incluídos no presente estudo os autores conseguiram determinar a mutação de sete deles segundo a metodologia proposta. Desses, três pacientes foram diagnosticados com mutações homozigoticas todas no gene CLRN1 e possuíam diagnostico clinico prévio de SU tipo 2. Dois pacientes apresentaram mutações heterozigóticas para o gene MYO7A, ambos com diagnostico clinico prévio de SU tipo 1 e um paciente apresentou mutação heterozigótica para o gene ALMS1 que apresentava diagnostico clinico de SU tipo 1.

La Retinosi Pigmentària (RP) és el terme aplicat a un grup de degeneracions de la retina clínicament i genèticament heterogeni. A nivell clínic es caracteritza per la presència d'una ceguesa nocturna i una constricció dels camps visuals, conduint a una pèrdua total de la visió. La RP pot heretar-se seguint la forma autosòmica dominant (RPAD), autosòmica recessiva (RPAR), i lligada al cromosoma X (RPLX), o seguint una forma d'herència digènica.Previs estudis han descrit mutacions en els gens PDEB, ABCA4, TULP1 i CNGA1 en un petit percentatge de famílies espanyoles amb RPAR. Aquestes dades indiquen que altres gens poden estar involucrats en la patologia de les restants famílies analitzades, emfatitzant l'elevada heterogeneïtat genètica present en aquesta malaltia i reforçant la hipòtesi que en la RPAR no hi ha un únic gen principal sinó que existirien varis gens on cadascun d'ells explicaria un petit número de casos. Hem analitzat la implicació de cinc gens addicionals que codifiquen per: la proteïna d'unió al retinaldehid (gen RLBP1), el receptor acoplat a proteïna G de l'EPR (gen RGR), la proteïna cel·lular d'unió al retinol (gen CRBP1), crumbs homologue 1 (gen CRB1) i la usherina (gen USH2A) en famílies espanyoles amb RPAR.Malgrat que en els gens RLBP1, RGR i CRBP1 s'han detectat diversos polimorfismes tant a nivell exònic com intrònic, no s'ha trobat cap mutació associada a la malaltia, suggerint que aquests gens probablement no estarien implicats en la patologia d'aquest grup de famílies espanyoles amb RPAR. En canvi, l'anàlisi mutacional dels gens CRB1 i USH2A ha permès la identificació de vàries variants rares, polimorfismes intrònics i diverses mutacions patològiques en el grup analitzat de pacients espanyols amb RPAR. L'ample rang de fenotips associats amb les mutacions trobades en els gens CRB1 i USH2A reflexa que les relacions entre les mutacions patològiques i el fenotip de la malaltia esdevenen cada cop més complexes.La síndrome d'Usher és la forma més freqüent de la RP sindròmica. Clínicament es caracteritza per una sordesa congènita i neurosensorial associada a RP. En un grup de pacients afectats amb la síndrome d'Usher tipus II es van detectar vàries mutacions patològiques en el gen USH2A. Els diferents fenotips associats amb les mutacions en el gen USH2A van revelar una important heterogeneïtat clínica en aquest grup de pacients. Aquests resultats junt amb els descrits en altres treballs confirmen una gran heterogeneïtat fenotipíca presentada per les mutacions en el gen USH2A, que va des de diferents tipus de la síndrome d'Usher fins a la RP no sindròmica. / Retinitis Pigmentosa (RP) is the term applied to a clinically and genetically heterogeneous group of retinal degeneration. Clinically is characterised by night blindness and constriction of visual fields, leading to complete blindness. RP can be inherited in either an autosomal dominant (ADRP), autosomal recessive (ARRP), X-linked (XLRP), or digenic mode. Previous studies have described mutations in the PDEB, ABCA4, TULP1 and CNGA1 genes in a small percentage of Spanish ARRP families. These data indicates that other than these genes may be involved in the remaining families analysed, emphasising the genetic heterogeneity of the disease and reinforcing the hypothesis that in ARRP there is no a major gene but several genes may account individually for a small number of cases.We analysed the involvement of five additional genes: the retinaldehyde-binding protein 1 (RLBP1 gene), the RPE retinal G protein-coupled receptor (RGR gene), the cellular retinol-binding protein 1 (CRBP1 gene), the crumbs homologue 1 (CRB1 gene) and usherin (USH2A gene) in ARRP Spanish families. Several exonic and intronic polymorphisms were detected in the RLBP1, RGR and CRBP1 genes. However, no disease-causing mutations were found, suggesting that these genes be most probably not involved in the disease in this set of ARRP Spanish pedigrees. In contrast, the mutational analysis of the CRB1 and USH2A genes allowed the identification of a number of rare sequence variants and intronic polymorphisms, and of several pathogenic mutations in the group of ARRP Spanish patients analysed. The wide range of phenotypes associated with mutations found in CRB1 and USH2A genes underlines how the relationship between pathogenic mutations and disease phenotype is becoming increasingly complex.The Usher syndrome is the most frequent syndromic RP. Clinically is characterised by congenital sensorineural hearing loss and RP. Several pathogenic mutations in USH2A gene were detected in a group of Usher syndrome type II patients. Different phenotypes associated with mutations in USH2A gene revealed an important heterogeneity clinical in this group of patients. The results presented here together with that in previous reports confirm a great phenotypic heterogeneity arising from mutations in USH2A gene, which ranges from distinct types of Usher syndrome to non-syndromic retinitis pigmentosa.

La (Pcdh15) est localisée dans les touffes ciliaires des cellules ciliées internes et externes de la cochlée. Elle forme des liens interstéréociliaires entre les différents stéréocils, des protrusions riches en actine qui forment ensemble la touffe ciliaire. L’absence de Pcdh15 entraîne la désorganisation des touffes ciliaires et la perte de la mécanotransduction auditive. Pcdh15 forme en effet la partie basse du lien de bout de cil (en anglais tip-link), le lien contrôlant l’ouverture du canal de transduction, qui se situe au point d’insertion inferieur de ce lien. Il existe trois isoformes de Pcdh15 (CD1, CD2 et CD3). J’ai étudié la distribution de ces isoformes dans la touffe ciliaire à différents stades de sa maturation. Différents modèles murins « KO conditionnel » ont été générés, ils m’ont permis d’analyser les conséquences de l’absence de chacune des isoformes individuellement, ainsi que celles de l’absence de Pcdh15-CD2 et Pcdh15-CD3, et celles de l’absence simultanée des trois isoformes. J’ai ainsi pu conclure que Pcdh15-CD2 est la seule isoforme essentielle pour la formation des tip-links dans les cellules ciliées matures. Dans les touffes ciliaires matures, Pcdh15 joue également un rôle dans l’interaction entre les touffes ciliaires et la membrane tectoriale, dans le contrôle de la taille des stéréocils, et dans la formation des liens interstéréociliaires apicaux. Pour ces fonctions, mais pas pour la formation du tip-link, Pcdh15-CD1 et Pcdh15-CD2 (les isoformes présentes dans la touffe ciliaire mature) sont redondantes. Cependant, Pcdh15-CD1 ne peut compenser que partiellement l’absence de Pcdh15-CD2 et Pcdh15-CD3. Pour étudier comment Pcdh15 interagit avec les autres protéines impliquées dans le syndrome de Usher, les interactions avec l’harmonine et la whirline (deux protéines d’échafaudage qui sont colocalisées avec Pcdh15 à l’apex des stéréocils) ont été analysées in vitro. / Protocadherin-15 (Pcdh15 is located in the stereociliary hair bundles of inner and outer hair cells (IHCs and OHCs) of the cochlea, where it forms fibrous links between different stereocilia. Absence of Pcdh15 leads to deafness due to the disorganization of hair bundles and absence of mechano-electrical transduction. The latter is explained as Pcdh15 forms the lower component of the tip-link, that gate hair cell mechano-electrical transduction channels. There are three different splice isoforms of Pcdh15 (CD1, CD2 and CD3), I studied their distribution in the developing and mature auditory hair cells. Different conditional Pcdh15 knockout mouse models were generated, permitting analysis of the absence of each of the different Pcdh15 isoforms individually, of the combined absence of Pcdh15-CD2 and Pcdh15-CD3, and of the absence of all isoforms. I was able to conclude that Pcdh15-CD2 is essential for the formation of tip-links in mature hair cells. In mature hair bundles Pcdh15 also plays a role in the coupling of the hair bundles to the tectorial membrane, in the control of the size of the stereocilia, and in the formation of apical links between stereocilia. The different Pcdh15 isoforms present in mature hair bundles (Pcdh15-CD1 and Pcdh15-CD2) are functionally redundant for these functions, but not for tip-link formation. In immature hair bundles, the different Pcdh15 isoforms are functionally redundant, although Pcdh15-CD1 can only partially compensate the absence of Pcdh15-CD2 and Pcdh15-CD3. To discover how Pcdh15 interacts with other proteins implicated in Usher syndrome, interactions with harmonin and whirlin were analyzed by biophysical techniques.

Visual conditions such as retinitis pigmentosa and Usher syndrome can gradually cause tunnel vision. Patients with these conditions usually face difficulties with navigation, avoiding obstacles, and performing visual search. Loss of mobility can affect patients' independence and quality of life. One of the rehabilitation strategies for patients with tunnel vision is the use of optical aids to enhance mobility performance. The main method used to evaluate the usefulness of optical aids is the patient’s subjective report after extended wear. In order to evaluate optical aid effectiveness in the clinic, a new test based on the visual search paradigm was designed to assess the patient's visual awareness. This was named the assessment of visual awareness (AVA) test. The main aim of this study was to develop the AVA test, establish its sensitivity, validity and repeatability, and then use it to investigate the efficacy of optical aids in this group of people. The AVA test consists of 32 peripheral targets presented at four different locations: 1st annulus (at 5° from the central fixation), 2nd annulus (10°), 3rd annulus (20°) and 4th annulus (30°)). In this study, the peripheral targets were presented singly against a spatial noise background in a presentation area of 81° H × 62° V. Participants were allowed to use head and eye movements and were asked to search for and locate each target. The detection time (DT) was recorded. A new, sensitive and easy to set up indoor mobility course was also designed and validated prior to its use in validating the AVA test. A total of 50 normally sighted participants with simulated tunnel vision (TV) (5° to 20°, in 5° steps) and 20 patients with TV (retained field 4° to 21°) were tested. The AVA test was found to be responsive to the change in field of view (FoV) and to the target locations in both groups of participants. In the simulated group, a significant relationship was found between FoV and DT at each annulus (r ranging from -0.55 to -0.77, p < 0.0001). A significant relationship was found between target location and DT within each FoV size (20°, 15°, 10° and 5°) (r ranging from 0.53 to 0.84, p < 0.0001). In the TV patients, a statistically significant relationship was found between FoV and DT at each annulus (r range from -0.40 to -0.60, p < 0.05). The target location was shown to have a significant relationship with the DT within each FoV size (r ranging from 0.50 to 0.60, p < 0.05). Finally, the AVA test was found to be significantly related to the simulated TV participants' performance on the indoor mobility course. The AVA test was used to assess the efficacy of three optical aids: the partial aperture prism (10 patients), the Tri-field prism (10 patients) and the reverse telescope (4 patients). The AVA test showed no significant improvement in DT with either of the prisms and the participants did not find these aids helpful. DT with the reverse telescope improved, but none of the participants were willing to use these on extended trial. The AVA test gave clear indications of the efficacy of each aid, a result which could affirm the importance of the AVA test. In conclusion, the AVA test was found to be sensitive, valid and repeatable. DT did not improve in either of the optical aids which were found to be unsuccessful, suggesting that the AVA could be a promising clinical test. However the aids which showed improved DT were not evaluated over the longer term, and therefore did not allow full evaluation of the AVA test.

Salmonella enterica sérovar Typhi est une bactérie pathogène humain-spécifique et l’agent étiologique de la fièvre typhoïde. Parmi ses facteurs de virulence, il y a 14 systèmes d’adhésion putatifs nommés fimbriae qui ont été identifiés dans le génome de S. Typhi. Les fimbriae sont regroupés en opérons qui codent pour des structures protéiques extracellulaires, pour une machinerie de sécrétion et d’assemblage et parfois pour des régulateurs. Ceux-ci sont peu exprimés en conditions de laboratoire et peu étudiés chez S. Typhi. Parmi les 14 fimbriae de S. Typhi, 12 appartiennent à la classe des chaperon-placier, c’est-à-dire qu’ils possèdent un chaperon et un placier qui leur sont dédiés pour la formation de la structure fimbriaire. Je crois que ces fimbriae sont importants pour la pathogenèse de S. Typhi. Le but de ce projet est l’analyse fonctionnelle des fimbriae de type chaperon-placier chez S. Typhi. Pour ce faire, j’ai voulu établir une caractérisation générale des 12 fimbriae de type chaperon-placier, puis j’ai concentré l’étude sur la régulation de 2 de ces fimbriae, c’est-à-dire Fim et Std. La caractérisation générale des fimbriae de type chaperon-placier consistait à déterminer l’expression des promoteurs fimbriaires lors de la croissance en différentes conditions de culture mimant l’infection, à déterminer la présence et la morphologie des fimbriae à la surface de la bactérie et à évaluer l’effet des fimbriae sur la pathogenèse de S. Typhi (formation de biofilm, interactions avec les cellules de l’hôte et motilité bactérienne). L’expression maximale des fimbriae a été obtenue principalement en milieu minimal. J’ai observé pour la première fois 6 des 12 fimbriae par microscopie électronique à transmission. Chaque fimbria présentait des effets sur au moins une étape testée sur la pathogénèse. La régulation de std et fim a été étudiée en déterminant le rôle de régulateurs globaux et par criblage d’une banque de mutants par insertion de transposon. Principalement, j’ai découvert que le promoteur std était activé par Crp, responsable de la répression catabolique, tandis que fim voit son expression modulée par la chaîne de transport d’électrons (Ndh) et des perturbations de l’enveloppe (OmpR). Finalement, nos résultats démontrent que les fimbriae de type chaperon-placier sont importants pour la 6 pathogenèse de S. Typhi et que deux de ceux-ci sont régulés par des signaux environnementaux importants rencontrés par la bactérie lors de l’infection. / Salmonella enterica serovar Typhi is a human-specific pathogenic bacteria and the etiologic agent of typhoid fever. Among its virulence factors, there are 14 putative adhesion systems named fimbriae identified in the S. Typhi genome. Each fimbria is clustered in an operon that encodes for extracellular proteinaceous structures, for the secretion and assembly machinery and sometime for regulators. Fimbrial genes are poorly expressed under laboratory conditions, with few studied in S. Typhi. Among the 14 fimbriae, 12 belong to the chaperone-usher class, where each one encodes a dedicated chaperone and usher that form the fimbrial structure. I propose that fimbriae are important for S. Typhi pathogenesis. The aim of this project is the functional analysis of all the chaperone-usher fimbriae of S. Typhi. My goals were to establish a general characterization of the 12 chaperone-usher fimbriae, and to study specifically the regulation of 2 fimbriae, Fim and Std. The general characterization of chaperone-usher fimbriae includes the determination of the expression of fimbrial promoters in different growth conditions mimicking infection, the observation of the presence and morphology of fimbriae at the bacterial surface, and the evaluation of the role of fimbriae on S. Typhi pathogenesis (biofilm formation, host-cells interactions and motility). Fimbrial expression was generally higher when cells were grown in minimal medium. I was able to observe for the first time the presence of 6 out of 12 fimbriae by transmission electron microscopy. Regarding the role of fimbriae in pathogenesis, each fimbria was involved in at least one step. Regulation of std and fim was studied by evaluating the implication of several general regulators and by screening a transposon-based library. Overall, I discovered that the std promoter was activated by Crp, responsible of catabolic repression, and that fim was modulated by the activity of the electron transport chain and by envelope perturbations. Finally, my results demonstrated that the chaperone-usher fimbriae are important for S. Typhi pathogenesis and two of them are regulated by important environmental signals encountered during bacterial infection.