Submitted by Renata Lopes (renatasil82@gmail.com) on 2017-04-06T19:57:29Z No. of bitstreams: 1 arnaldofernandescorrea.pdf: 1337257 bytes, checksum: 10475519e434ae4106974b9bcb37665c (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-04-07T16:11:39Z (GMT) No. of bitstreams: 1 arnaldofernandescorrea.pdf: 1337257 bytes, checksum: 10475519e434ae4106974b9bcb37665c (MD5) / Made available in DSpace on 2017-04-07T16:11:39Z (GMT). No. of bitstreams: 1 arnaldofernandescorrea.pdf: 1337257 bytes, checksum: 10475519e434ae4106974b9bcb37665c (MD5) Previous issue date: 2014-08-12 / A proposta que as instituições públicas contemporâneas almejam é que os Estados brasileiros se transformem e construam no interior destas, ambientes de trabalho mais colaborativos, integrados, arrojados e técnicos. Esta dissertação pretende explicar o processo de implementação da Diretoria de Pessoal (DIPE) na Superintendência Regional de Ensino de Ubá (SRE/Ubá) que conforme a nova estrutura orgânica da Secretaria de Estado de Educação de Minas Gerais (SEE/MG) passou a vigorar a partir de 27/01/2011, mediante o Decreto nº 45.536, em todas as SREs. A pesquisa visa a contribuir para o entendimento da seguinte questão: A nova estrutura organizacional implementada na SRE/Ubá proporciona qualidade na prestação de serviço ao público em geral? Desse modo, o objetivo geral é analisar se esta nova estrutura está promovendo trabalhos alinhados às demandas do público que procura a SRE/Ubá. Os específicos focam em investigar se tal mudança proporciona cooperação estratégica governamental, a partir de melhor organização das funções, ao orientar os setores na busca de soluções eficazes para o alcance das metas pactuadas. Justifica-se o tema, uma vez que a DIPE caminha para superar dificuldades, apoiar e qualificar os servidores que atuam nesta área, tendo como foco a melhoria da qualidade dos serviços ofertados. A metodologia envolveu um Estudo de Caso cuja pesquisa de campo foi exploratória, descritiva e de cunho qualitativo, tendo como técnicas os seguintes instrumentos utilizados: questionário, entrevista com roteiro semiestruturado e análise documental tais como: leis, decretos e notas técnicas, além dos conceitos de alguns teóricos, a saber: Chiavenato, Bergue, Tachizawa, Paradela e Mello. A partir de alguns resultados apresentados, a DIPE proporcionou maior eficiência na execução dos trabalhos, bem como na assistência ao público, promovendo maior diálogo entre SRE-Escola, com consequente descentralização das tarefas e facilidade de comunicação, tornando assim, o serviço mais qualificado e ágil. / The proposal that contemporary public institutions crave is that Brazilian states to transform and build within them, work environments more collaborative, integrated, and daring technicians. This dissertation aims to explain the implementation process of the Directorate of Personnel (DIPE) in the Regional Superintendent of Education of Uba (SRE / Uba) that according to the new organizational structure of the State Department of Education of Minas Gerais (SEE / MG) became effective from 27/01/2011, by Decree No. 45,536, in all SREs. The research aims to contribute to understanding the following question: The new organizational structure implemented in SRE / Uba provides quality in providing service to the general public? Thus, the general objective is to analyze whether this new structure is promoting work aligned to the demands of the public who seeks SRE / Uba. The specific focus on investigating whether such a change provides strategic government cooperation, from better organization of functions, to guide the sector in seeking effective solutions to achieve the agreed targets. Justified the theme, since the DIPE walks to overcome difficulties, support and qualify the servers who work in this area, focusing on improving the quality of services offered. The methodology involved a case study whose field research was exploratory, descriptive and qualitative nature, with the techniques used the following instruments: questionnaire, semistructured interview and document analysis such as laws, decrees and technical notes, beyond concepts some theorists, namely: Chiavenato, Bergue, Tachizawa, Paradela and Mello. From some results presented in DIPE provided greater efficiency in the execution of the works, as well as assistance to the public, promoting greater dialogue between SRE-School, with consequent decentralization of tasks and ease of communication, thus making the most qualified service and agile.

Identification of novel cancer driver mutations is crucial for targeted cancer therapy, yet a difficult task especially for low frequency drivers. To identify cancer driver mutations, we developed a machine learning (ML) model to predict cancer hotspots. Here, we applied the ML program to 32 non-receptor tyrosine kinases (NRTKs) and identified 36 potential cancer driver mutations, with high probability mutations in 10 genes, including ABL1, ABL2, JAK1, JAK3, and ACK1. ACK1 is a member of the poorly understood ACK family of NRTKs that also includes TNK1. Although ACK1 is an established oncogene and high-interest therapeutic target, the exact mechanism of ACK1 regulation is largely unknown and there is still no ACK1 inhibitor in clinical use. The ACK kinase family has a unique domain arrangement with most notably, a predicted ubiquitin association (UBA) domain at its C-terminus. While the presence of a functional UBA domain on a kinase is unique to the ACK family, but the role of the UBA domain on ACK1 is unknown. Interestingly, the ML program identified the ACK1 Mig6 homology region (MHR) and UBA domains truncating mutation p633fs* as a cancer driver mutation. Our data suggest that the ACK1 UBA domain helps activate full-length ACK1 through induced proximity. It also acts as a mechanism of negative feedback by tethering ACK1 to ubiquitinated cargo that is ultimately degraded. Indeed, our preliminary data suggest that truncation of the ACK1 UBA stabilizes ACK1 protein levels, which results in spontaneous ACK1 oligomerization and activation. Furthermore, our data suggests removal of the MHR domain hyper activates ACK1. Thus, our data provide a model to explain how human mutations in ACK1 convert the kinase into an oncogenic driver. In conclusion, our data reveal a mechanism of ACK1 activation and potential strategies to target the kinase in cancer.

Structuraj investigations on the Ddi1 (DNA-damage inducible protein 1) of Leishmania major and on the rotavirus nonstructural protein NSP4 were carried out. Ddi1 belongs to the ubiquitin receptor family of proteins. One of its domains is similar to the retroviral aspartic proteinases. It has been shown that this domain is the target of HIV-protease inhibitors that were being used in the treatment of AIDS and it was observed that these drugs effectively controlled opportunistic diseases caused by many parasitic protozoa such as Leishmania and Plasmodium species. The retroviral protease-like domains present in Ddi1 proteins of these organisms were identified as the targets of these drugs. Structural studies on Ddi1 from L. major have been carried out, in an attempt to provide a platform for the design of anti-protozoal compounds. Rotavirus NSP4, the first viral enterotoxin to be identified, is a multifunctional glycoprotein that plays critical roles in viral pathogenesis and morphogenesis. As part of an ongoing project on the structural characterization of NSP4, we determined the structure of the diarrhea-inducing region of this protein from the rotavirus strain MF66. Chapter 1 presents an overview of Ddi1 and NSP4 of the rotavirus with an emphasis on their structural features. The methods employed during the course of the present work are described in Chapter 2. Structural studies on the retroviral protease-like domain of Ddi1 (Ddi1-RVP) of L. major is presented in Chapter 3. Apart from this domain, Ddi1 of L. major also has a ubiquitin-associated and ubiquitin-like domains whereas P. falciparum has only the ubiquitin-associated domain. Activity of the full length Ddi1 of L. major and the retroviral protease domain of P. falciparum using an HIV protease substrate was shown to be inhibited by an HIV protease inhibitor, saquinavir. Binding of saquinavir to the proteins was also confirmed by Biolayer Interferometry studies. The crystal structure of the retroviral protease domain of L. major Ddi1 has been determined. It forms a homodimeric structure similar to that of HIV protease and the reported structure of the same domain from Saccharomyces cerevisiae. The loops in Ddi1-RVP are similar to the 'flap' regions of the HIV protease which close-in upon substrate/inhibitor binding; they are visible in the electron density maps, unlike the case of the S. cerevisiae protein. Though the native form of the domain shows an open dimeric structure, normal mode analysis reveals that it can take up a closed conformation resulting from relative movements of the subunits. The present structure of Ddi1-RVP of L. major with the defined 'flap'-like loops will be helpful in the design of effective drugs against protozoal diseases, starting with HIV protease inhibitors as the lead compounds. Chapter 4 describes the structural investigations carried out on the diarrhea-inducing region of the nonstructural protein NSP4 of the rotavirus strain MF66 which forms an α-helical coiled-coil structure. Crystal structures of a synthetic peptide and of two recombinant proteins spanning this region showed parallel tetrameric organization of this domain with a bound Ca2+ ion at the core. Subsequently, we determined the structure of NSP4 from a different strain as a pentamer without the bound Ca2+ ion. This new structure provides more insights into understanding some of the functions of NSP4 such as the release of ions into the cytoplasm and binding to the double-layered particle (DLP). We also established conditions responsible for these structural transitions. The crystal structure of the coiled-coil domain of NSP4 presented in this chapter shows an entirely different structure which is an antiparallel tetramer. This explains our failure to determine the structure by the molecular replacement method using known oligomers. The structure was solved by the Sulphur-SAD method using diffraction data collected with Cr Ka radiation. The study reveals that the structural diversity of NSP4 is not limited. We could relate sequence variations and pH conditions to the differences in oligomeric assemblies. Surface properties of the domain suggest that the new form is likely to interact with different sets of proteins compared to those that interact with the parallel tetramers or pentamers. Further investigations are needed to establish this property.