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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2006.tde-03122021-150906
Document
Author
Full name
Natalia Pasternak Taschner
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2006
Supervisor
Committee
Spira, Beny (President)
Baldini, Regina Lúcia
Marques, Marilis do Valle
Menck, Carlos Frederico Martins
Silva, Aline Maria da
Title in Portuguese
A regulação da fosfatase alcalina pelo fator σS da RNA polimerase de Escherichia coli.
Keywords in Portuguese
fator sigma
fosfatase alcalina
operon pst
regulon PHO
RNA polimerase
Abstract in Portuguese
O fator σS associado à RNA polimerase de E. coli (EσS) é responsável pela transcrição da maioria dos genes relacionados à sobrevivência durante a fase estacionária e em períodos de estresse. Quando a célula entra em carência de fosfato inorgânico, um conjunto de genes e operons conhecidos como o regulon PHO é ativado. Os genes do regulon PHO são transcritos preferencialmente pela RNA polimerase associada ao fator σD. A inativação do gene rpoS, que codifica para o fator σS, causou um aumento significativo na expressão da maioria dos genes de PHO. O efeito repressor de σS foi atribuído à competição entre este fator sigma e o fator σD pelo cerne da RNA polimerase. Diferentemente dos demais genes de PHO, a expressão de pstS, o primeiro gene do operon pst, foi moderadamente estimulada na presença do fator σS. A região promotora de pstS contém uma citosina na posição -13 e um sítio de ligação para a proteína IHF. Estes elementos, encontrados em genes dependentes do fator σS, podem estar contribuindo para a transcrição de pstS pela holoenzima EσS. Um estudo do polimorfismo do gene rpoS em diversas cepas de E. coli K-12 detectou a presença de uma mutação âmbar no códon 33 deste gene. Bactérias supressoras da mutação âmbar produzem uma proteína de tamanho normal (38 KDa), ao passo que bactérias não-supressoras produzem um fator σS truncado, que apesar de possuir 55 aminoácidos a menos, foi capaz de transcrever seus genes dependentes ainda que com menor eficiência. Cepas que carregam o códon 33 âmbar apresentaram um fenótipo diferenciado em relação à expressão dos genes de PHO.
Title in English
Alkaline phosphatase regulation by the σS factor of RNA polymerase in Escherichia coli.
Keywords in English
pst operon
alkaline phosphatase
PHO regulon
RNA polymerase
sigma factor
Abstract in English
In E. coli, the RNA polymerase associated with σS (EσS) is responsible for the transcription of the majority of the genes related to survival during stress periods or during the stationary phase of growth. When cells enter a phase of phosphate starvation, the transcription of several genes and operons, known as the PHO regulon, is activated. Knock-out of rpoS, which encodes σS, increases the expression of most PHO genes. The negative effect of σS on PHO expression is probably due to the competition between σS and σD for the core RNA polymerase. Unlike all other PHO genes, pstS, the first gene of the pst operon, was mildly stimulated by σS. The promoter region of pstS contains a cytosine residue at position -13, and an IHF binding site. These elements are known to be important for the transcription of σS-dependent genes, and may contribute for the transcription of pstS by EσS . The polymorphism of rpoS in several E. coli K-12 strains was evaluated. An amber mutation in codon 33 of this gene was found in 2 out of 7 strains. These strains were found to suppress the amber mutation and to produce a normal 38 KDa σS protein. Non-suppressor strains carrying this amber mutation produced a σS variant, lacking 55 amino acids. The truncated σS protein was still able to drive the transcription of its dependent genes, albeit in a smaller proportion. Strains carrying the rpoS amber mutation presented a distinct phenotype regarding the expression of the PHO regulon genes.
 
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Publishing Date
2021-12-06
 
WARNING: The material described below relates to works resulting from this thesis or dissertation. The contents of these works are the author's responsibility.
  • TASCHNER, N. P., and SPIRA, B. A differential effect of S on the expression of the PHO regulon genes of Escherichia coli [doi:10.1099/mic.0.27124-0]. Microbiology (Reading) [online], 2004, vol. 150, nº 9, p. 2985-2992.
  • Taschner, Natalia Pasternak, Yagil, Ezra, and Spira, Beny. The effect of IHF on σ S selectivity of the phoA and pst promoters of Escherichia coli [doi:10.1007/s00203-005-0082-4]. Archives of Microbiology [online], 2006, vol. 185, nº 3, p. 234-237.
  • TASCHNER, N. P., e SPIRA, B. Regulation of alakaline phosphatase by the sigmaS factor of RNA polymerase in Escherichia coli. In XXIII Reunião de Genetica de Microrganismos, Pirenopolis, 2002. XXIII Reunião de Genetica de Microrganismos., 2002. Resumo.
  • Premio "melhor trabalho publicado" pela aluna de pós-graduação Natalia Taschner - IV Reunião Científica do Departamento de Microbiologia
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