Leonardo Talachia Rosa

Pós-doutorado

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Áreas de atuação

Participação em eventos

Participação em bancas

Orientou

Produções bibliográficas

• JUR'NAS, DUKAS ; ROSA, LEONARDO TALACHIA ; REY, MARTIAL ; CHAMOT-ROOKE, JULIA ; FRONZES, RÉMI ; CASCALES, ERIC . Mounting, structure and autocleavage of a type VI secretion-associated Rhs polymorphic toxin. Nature Communications , v. 12, p. 6998, 2021.

• ROSA, LEONARDO T. ; SPRINGTHORPE, VICKI ; BIANCONI, MATHEUS E. ; THOMAS, GAVIN H. ; KELLY, DAVID J. . Massive over-representation of solute-binding proteins (SBPs) from the tripartite tricarboxylate transporter (TTT) family in the genome of the -proteobacterium Rhodoplanes sp. Z2-YC6860. Microbial Genomics , v. 4, p. 176, 2018.

• ROSA, LEONARDO T. ; DIX, SAMUEL R. ; RAFFERTY, JOHN B. ; KELLY, DAVID J. . A New Mechanism for High-Affinity Uptake of C4-Dicarboxylates in Bacteria Revealed by the Structure of Rhodopseudomonas palustris MatC (RPA3494), a Periplasmic Binding-Protein of the Tripartite Tricarboxylate Transporter (TTT) Family. JOURNAL OF MOLECULAR BIOLOGY , v. 1, p. YJMBI 65926, 2018.

• ROSA, LEONARDO T. ; BIANCONI, MATHEUS E. ; THOMAS, GAVIN H. ; KELLY, DAVID J. . Tripartite ATP-Independent Periplasmic (TRAP) Transporters and Tripartite Tricarboxylate Transporters (TTT): From Uptake to Pathogenicity. Frontiers in Cellular and Infection Microbiology , v. 8, p. 324356, 2018.

• ROSA, LEONARDO T ; DIX, SAMUEL R ; RAFFERTY, JOHN B ; KELLY, DAVID J . Structural basis for high-affinity adipate binding to AdpC (RPA4515), an orphan periplasmic binding protein from the Tripartite Tricarboxylate Transporter (TTT) family in. FEBS Journal , v. -, p. 14304, 2017.

• ROSA, LEONARDO TALACHIA ; RAFFERTY, JOHN ; KELLY, DAVID . Biotechnological potential of novel TTT-family transporters from Rhodopseudomonas palustris. New Biotechnology , v. 33, p. S37, 2016.

• ROSA, L. T. ; VERNHES, E. ; POLARD, P. ; Fronzes, R . showing the way: RadA helicase mechanism deciphered by CryoEM. 2023. (Apresentação de Trabalho/Comunicação).

• ROSA, L. T. ; VERNHES, E. ; POLARD, P. ; FRONZES, RÉMI . Showing the way: RadA helicase mechanism during natural transformation in Streptococcus pneumoniae shown by CryoEM. 2022. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. . Uso de biologia estrutural para elaboração de modelos de mecanismos biológicos. 2022. (Apresentação de Trabalho/Conferência ou palestra).

• ROSA, L. T. ; VERNHES, E. ; POLARD, P. ; Fronzes, R . Structural Aspects of the Transformation-Dedicated Helicase RadA from S. Pneumoniae. 2021. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. ; VERNHES, E. ; POLARD, P. ; Fronzes, R . CryoEM structure of RadA, a Superfamily 4 modular helicase involved in natural transformation in Streptococcus pneumoniae. 2021. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . Biotechnological potential of novel TTT-family transporters from Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. . Metabolite transporters and aromatic compound utilisation by the photosynthetic bacteriaRhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, DAVID . high-affinity adipate binding to AdpC , an orphan periplasmic binding protein from the Tripartite Tricarboxylate Transporter (TTT) family in Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. ; Dixon, S ; RAFFERTY, JOHN ; KELLY, D. J. . Biotechnological potential of novel TTT-family transporters from Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Seminário).

• ROSA, L. T. ; Dixon, S ; RAFFERTY, JOHN ; KELLY, D. J. . high-affinity adipate binding to AdpC an orphan periplasmic binding protein from the Tripartite Tricarboxylate Transporter (TTT) family in Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. . Bacterial Insight - The faster you chew, the more you eat. 2017. (Apresentação de Trabalho/Seminário).

• ROSA, L. T. . TTT transporters in Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, LEONARDO TALACHIA ; RAFFERTY, JOHN ; KELLY, DAVID ; Dixon, S . Structural basis for high-affinity adipate binding to AdpC (RPA4515), an orphan periplasmic binding protein from the Tripartite Tricarboxylate Transporter (TTT) family in Rhodopseudomonas palustris. 2017. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . TTT transporters in Rhodopseudomonas palustris. 2016. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . Biotechnological potential of novel TTT-family transporters from Rhodopseudomonas palustris. 2016. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. . Lignin degradadion in Rhodopseudomonas palustris. 2016. (Apresentação de Trabalho/Simpósio).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . TTT transporters in Rhodopseudomonas palustris. 2016. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . Biotechnological potential of TTT transporters in Rhodopseudomonas palustris. 2016. (Apresentação de Trabalho/Conferência ou palestra).

• ROSA, L. T. . Microbiology in biotechnology and synthetic biology. 2016. (Apresentação de Trabalho/Conferência ou palestra).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . TTT transporters in Rhodopseudomonas palustris. 2015. (Apresentação de Trabalho/Conferência ou palestra).

• ROSA, L. T. ; RAFFERTY, J. B. ; KELLY, D. J. . Characterization of secondary soluble binding protein dependent transporters in Rhodopseudomonas palustris. 2015. (Apresentação de Trabalho/Conferência ou palestra).

• ROSA, L. T. ; Rodrigues, Edson ; Trapp, Marilia A. . PROSPECÇÃO DE COMPOSTOS ANTIBIÓTICOS PRODUZIDOS POR BACTÉRIAS ENDOFÍTICAS ASSOCIADAS À PLANTA Alternanthera brasiliana (Amaranthaceae). 2011. (Apresentação de Trabalho/Congresso).

• Rodrigues, Edson ; Trapp, Marilia A. ; ROSA, L. T. . Identification of endophytic bacteria from Alternanthera brasiliana by MALDI-TOF. 2011. (Apresentação de Trabalho/Congresso).

• ROSA, L. T. ; Malavazi, I. . Caracterização parcial do gene que codifica uma proteína quinase C (PKCA) no fungo patogênico Aspergillus fumigatus. 2010. (Apresentação de Trabalho/Congresso).

Outras produções

Projetos de pesquisa

• 2023 - Atual

  The Tripartite Tricarboxylate Transporter (TTT) family: Investigating signalling and transport networks in the re-emerging pathogen Bordetella pertussis, Descrição: Pertussis, or whooping cough, is a pulmonary-tract disease caused by the Gram-negative bacterium Bordetella pertussis. Affecting mainly children under four years old, this disease is characterized by persistent cycles of cough which can lead to pulmonary tissue damage and necrosis. Despite the existing vaccine, several new pertussis epidemies have been described over the last years, elevating the status of B. pertussis to re-emerging pathogen. Encoded on the genome of species from the Bordetella genus, a family of high affinity uptake systems, the Tripartite Tricarboxylate Transporters (TTT) is found to be highly overrepresented. The TTT family is composed by a 12-15 transmembrane translocator domain TctA, a four transmembrane domain with unknown function TctB and a periplasmic Substrate Binding Protein (SBP) TctC. B. pertussis have 78 genes encoding for TctC proteins, making it the most abundant gene family in the genome, in contrast to only 2 TctAB systems. Widespread in the bacterial kingdom, the TTT family is particularly abundant in alpha and beta-proteobacteria. However, B. pertussis is the only strict pathogen to show a similar expansion. Despite initial indications pointing to an involvement of the TTT SBPs in virulence and core metabolism, the role of this protein family in B. pertussis remains elusive. Moreover, the ligand translocation mechanism of the TTT family remains unknown. This study aims to (i) determine the translocation mechanism of the TTT family studying the citrate transport system TctCBA as a prototype, using CryoEM, (ii) investigate the involvement of the TctC homologs in the core metabolism and pathogenicity of this reemeging pathogen and (iii) design a molecular inhibitor of the TctC homologs in order to collapse B. pertussis metabolism. Together, these results will deepen our understanding of B. pertussis metabolism and pathogenesis process, leading to potential novel drug targets for the treatment of whooping cough, as well as elucidate the transport mechanism of a family of transporters ubiquitously found among bacteria.. , Situação: Em andamento; Natureza: Pesquisa. , Integrantes: Leonardo Talachia Rosa - Coordenador., Financiador(es): Fundação de Amparo à Pesquisa do Estado de São Paulo - Auxílio financeiro.

• 2023 - Atual

  CEPID-B3 - Biology of Bacteria and Bacteriophages Research Center, Descrição: The CEPID B3 is a network with research hubs based in São Paulo (IQ-USP, ICB-USP), Campinas (IB-UNICAMP, IAC), Ribeirão Preto (FCFRP-USP, FMRP-USP) and Rio Claro (UNESP) that will collaborate to carry out cutting edge basic research into molecular mechanisms that are of prime importance to understand bacterial reproduction, multicellular behavior, competition, colonization and interactions with their eukaryotic hosts and bacteriophages. The acquired knowledge will be used in the search and development of new chemical and biological inhibitors of bacterial growth.. , Situação: Em andamento; Natureza: Pesquisa. , Integrantes: Leonardo Talachia Rosa - Integrante / Shaker Chuck Farah - Coordenador., Financiador(es): Fundação de Amparo à Pesquisa do Estado de São Paulo - Auxílio financeiro.

• 2022 - 2023

  Sistemas de secreção do tipo IV conjugativos na família Xanthomonadaceae: Investigando diversidade estrutural e funcional, Descrição: Type IV Secretion systems (T4SS) are macromolecular protein complexes in bacteria which, encompassing the membranes, mediate the exchange of DNA and protein effectors to target cells, playing a key role in conjugation, bacterial competition, eukaryotic cell invasion and immune system evasion in different species. In the last fifteen years, our group have been studying the role of bacteria-killing T4SS in Xanthomonas citri, the causative agent of canker citrus, and in the opportunistic pathogen Stenotrophomonas maltophilia, both belonging to the Xanthomonadaceae family. Our genome searches revealed that these two species contain each an additional T4SS cluster, so far uncharacterized and annotated as participating in bacterial conjugation. These systems have in common the absence of a VirB7 homolog, a so far universal component of the Outer Membrane Core Complex (OMCC), providing a convenient model to explore the minimum requirements for T4SS assembly. Moreover, the genomic location and distance from conjugative components in the S. maltophilia system indicate that its primary function differs from bacterial conjugation, and we hypothesize it may participate in eukaryotic cell invasion in the context of opportunistic pathogenicity. Thus, we aim to evaluate the participation of these two T4SS in conjugation and eukaryotic cell invasion, as well as to define the structure of their OMCC using Single-Particle Analysis Cryo-electronic microscopy (SPA-CryoEM). Minor components of these systems with unknown function will also be studied at their physiological and structural level. The results generated by this project will expand the knowledge on the function and structural diversity of T4SS, as well as on the metabolism of free-living and opportunistic pathogenic bacteria.. , Situação: Concluído; Natureza: Pesquisa. , Integrantes: Leonardo Talachia Rosa - Integrante / Shaker Chuck Farah - Coordenador., Financiador(es): Fundação de Amparo à Pesquisa do Estado de São Paulo - Bolsa.

• 2018 - 2022

  Structural study of the transformassome machinery in Gram positive bacteria using CryoEM, Descrição: Natural genetic transformation, first discovered in Streptococcus pneumoniae by Griffith in 1928, is observed in many Gram-negative and Gram-positive bacteria. This process promotes genome plasticity and adaptability. In particular, it enables many human pathogens such as Streptococcus pneumoniae, Staphylococcus aureus or Neisseria gonorrhoeae to acquire resistance to antibiotics and/or to escape vaccines. For natural transformation to occur, bacteria must develop a highly regulated physiological state called competence, during which a multiprotein machinery, designated as the transformasome, is specifically expressed. The transformasome drives the uptake of extracellular DNA and its subsequent integration into the bacterial genome. This intricate system can be divided into three functional entities: the transformation pilus, the DNA entry pore and the recombination apparatus. The architecture and function of the transformasome remains largely elusive. We propose to dissect the structure and function of this system in the human pathogen Streptococcus pneumoniae. We will combine four complementary approaches to attain our goal: (1) Molecular biology and biochemical studies in vitro (2) Structural biology on purified proteins or protein complexes using X-ray crystallography and cryo-electron microscopy (3) Structure-function studies in vitro and in vivo (4) Structural biology in situ using cryo-tomography. Overall, we believe this ambitious project will provide major insights to understand the molecular basis of bacterial transformation, a fundamental process in bacterial physiology. In addition, this project may provide new strategies to reduce pneumococcal adaptation during infection.. , Situação: Concluído; Natureza: Pesquisa. , Alunos envolvidos: Especialização: (1) Doutorado: (2) . , Integrantes: Leonardo Talachia Rosa - Integrante / Remi Fronzes - Coordenador / Robin ANGER - Integrante / Thomas Perry - Integrante / Esther Gavello Fernandes - Integrante.

• 2014 - 2018

  The Tripartite Tricarboxylate Transporters (TTT) : The neglected family of high-affinity uptake systems, Descrição: Bacteria inhabiting complex environments must possess efficient ways of acquiring nutrients, often present in low concentrations. In this context, Solute Binding Protein (SBP) dependent transporters make use of a periplasmic binding protein to bind substrates with high affinity, and deliver it to their transmembrane counterparts. There are three known families of SBP dependent transporters: The ABC (ATP-Binding Cassette), the TRAP (TRipartite ATP-independent Periplasmic transporters) and the TTT (Tripartite Tricarboxylate Transporter). The latter are very poorly characterized and only a few types of substrates for TTT transporters are currently known. This thesis first presents a review of the TTT transporters. Our database searches reveal a massive overrepresentation of TTT SBPs among and -proteobacteria, and highlight the presence of 434 TTT SBPs in the bacterium Rhodoplanes sp. Z2-YC6860, the biggest gene family representation described in a bacterial genome to date. We subsequently focus on the characterization of the TTT family in Rhodopseudomonas palustris, a model soil non-sulfur purple bacterium. The TTT family in R. palustris is formed by two complete tripartite systems, plus five orphan SBPs with no obvious membrane counterparts. From the seven SBPs, we present high-resolution crystal structures for six of them, and two of them were characterized biochemically and physiologically. One of the orphan SBPs (AdpC) is described to bind to dicarboxylic acids ranging from six to nine carbons in length with low M affinity, and is more expressed under low concentration of adipate. The MatC protein, belonging to the tripartite system MatBAC, is shown to bind to malate with low nM affinity. This study also attempted to provide the first biochemical characterization of a TTT tripartite system, with the reconstitution of the transmembrane components MatBA into proteoliposomes. In summary, this study presents a characterization of the Tripartite Tricarboxylate Transporter family in bacteria, through a synergistic multidisciplinary approach.. , Situação: Concluído; Natureza: Pesquisa. , Alunos envolvidos: Doutorado: (2) . , Integrantes: Leonardo Talachia Rosa - Integrante / David J Kelly - Coordenador., Financiador(es): Conselho Nacional de Desenvolvimento Científico e Tecnológico - Bolsa., Número de produções C, T & A: 3

• 2012 - 2013

  Knock-out of TRAP and ABC transport system involved with aromatic compounds uptake in Rhodopseudomonas palustris, Descrição: Aromatic compounds represent an industrial problem in varied situations. Although being usually toxic for most bacteria, some organisms were found to uptake and degrade these substances only with their own metabolic machinery. One of the most promising organisms, in this context, is Rhodopseudomonas palustris. The Cou operon contains the first genes involved in coumarate degradation. Within the Cou operon, there are two transport systems, showing that the uptake of aromatic compounds is co-regulated with their degradation. Curiously, the two transport systems works by different mechanisms. RPA1782-1784 is predicted to encode a TRAP (Tripartite ATP-independent periplasmic) transporter, a group of secondary transporters which uses the proton-motive force (pmf) as source of energy, and, in opposite of most secondary transporters, has high affinity for the substrate, mediated by a high-affinity periplasmic binding protein. This complex was further named TarPQM (TRAP transporter for aromatic compounds). On the other hand, RPA1789/RPA1791-1793 encodes for a classical ABC transporter, a primary transport system, thus using ATP as energy source. Despite having similar folding, the two binding proteins show insignificant sequence similarities, representing then a probable event of convergent evolution. Such event reinforces the importance of aromatic compounds uptake in soil environment, where competition is high and substrate availability is not. In this context, this study aims to produce null mutants for these transporters, to further characterize their role.. , Situação: Concluído; Natureza: Pesquisa. , Alunos envolvidos: Graduação: (1) / Doutorado: (2) . , Integrantes: Leonardo Talachia Rosa - Integrante / David J Kelly - Coordenador / Robert Salmon - Integrante., Financiador(es): Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Bolsa.

• 2011 - 2012

  Identificação de bactérias endofíticas associadas à planta Alternanthera brasiliana (Amaranthaceae) e prospecção de seus compostosantibióticos, Descrição: The demand for new antimicrobial compounds increases in our society. The resistance profile of pathogenic organisms to the available antibiotics is concerning, where relates of multidrug resistant organism are widespread. Therefore, the resumption of searches for new compounds from natural sources is needed, allied to design techniques. In this context, this work aimed to find, through dereplication methodology, antibiotic compounds produced by endophytic bacteria associated to the plant Alternanthera brasiliana, known by its antibiotic properties in popular medicine. Thus, 32 bacteria were isolated from plant tissues, where one of them, Bacillus amyloliquefaciens, showed itself as a producer of an antibiotic molecule, of m/z 803, possibly belonging to the group of depsipeptides. The study shows, moreover, the potential of mass spectrometry MALDI-TOF to the identification of endophytic microorganisms.. , Situação: Concluído; Natureza: Pesquisa. , Alunos envolvidos: Graduação: (1) / Doutorado: (2) . , Integrantes: Leonardo Talachia Rosa - Integrante / Edson rodrigues filho - Coordenador / Marilia Trapp - Integrante., Financiador(es): Fundação de Amparo à Pesquisa do Estado de SP - Bolsa.

• 2009 - 2010

  Isolamento de linhagens mutantes e caracterização parcial do gene que codifica uma proteína quinase C (PkcA) no fungo patogênico humano Aspergillus fumigatus, Descrição: O fungo filamentoso Aspergillus fumigatus, normalmente presente em material em decomposição, encontra no aparelho respiratório de indivíduos imunocomprometidos um local favorável ao seu estabelecimento, desencadeando o quadro de aspergillose invasiva à medida que coloniza o pulmão do hospedeiro. Essa doença representa uma das maiores causas de infecção hospitalar, apresentando taxa de mortalidade superior a 50%. Para que possa sobreviver no hospedeiro mamífero, o fungo deve adaptar-se aos estresses diversos encontrados no sítio de infecção, o que inclui, por exemplo, diferenças na pressão de oxigênio, perfil nutricional e exposição a espécies reativas de oxigênio. Dentre os mecanismos envolvidos nessa adaptação, a via mediada pela proteína quinase C (PkcA) merece destaque por estar diretamente envolvida na manutenção e reestruturação da parede celular. A resposta mediada pela Pkc envolve a ativação ?downstrean? de cascata de MAP quinases e em última instância a regulação de cerca de 25 genes envolvidos no processo de adaptação. Nosso objetivo é a caracterização dessa via metabólica no fungo patógeno humano A. fumigatus e sua relação patogênese e virulência.. , Situação: Concluído; Natureza: Pesquisa. , Alunos envolvidos: Graduação: (1) / Doutorado: (1) . , Integrantes: Leonardo Talachia Rosa - Integrante / Iran Malavazi - Coordenador., Financiador(es): Universidade Federal de São Carlos - Bolsa.

Prêmios