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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2012.tde-09022012-162000
Document
Author
Full name
Juan Carlos Guerrero Abad
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2011
Supervisor
Committee
Moura, Daniel Scherer de (President)
Brandão, Marcelo Mendes
Nogueira, Fabio Tebaldi Silveira
Title in Portuguese
Identificação da subtilase responsável pelo processamento do prepopeptídeo AtRALF1 em Arabidopsis thaliana
Keywords in Portuguese
Desenvolvimento vegetal
Enzimas - Identificação
Expressão gênica
Peptídeos hormonais
Proteínas de plantas
Abstract in Portuguese
Desde a década de 90, uma nova família de moléculas de origem protéica e com características hormonais vem sendo estudada em plantas. Esse grupo de novas moléculas, coletivamente chamado de peptídeos hormonais, está envolvido com defesa, reprodução, crescimento e desenvolvimento. RALF, que é um desses novos peptídeos, é ubíquo em plantas e está envolvido com desenvolvimento vegetal. Em Arabidopsis existem 34 genes semelhantes ao RALF (AtRALFs). Nosso grupo mostrou que AtRALF1 é processado de um precursor maior por uma subtilase. Arabidopsis possui 56 subtilases, o objetivo deste trabalho é identificar a subtilase responsável pelo processamento do AtRALF1. Predição da localização subcelular e análise da expressão in silico, ambas confirmadas por análise da expressão por RT-PCR e fusões proteicas com a proteína verde fluorescente, permitiram reduzir para sete o número de subtilases candidatas. Cruzamentos entre mutantes nocaute ou plantas de baixa expressão por RNAi dessas sete subtilases com plantas superexpressoras do AtRALF1 identificaram as subtilases AtSBT6.1 (At5g19660) e AtSBT5.3 (At2g04160) como prováveis envolvidas no processamento do prepropeptídeo AtRALF1.
Title in English
Identification of the subtilase responsible for the prepropeptide AtRALF1 processing in Arabidopsis thaliana
Keywords in English
Enzymes -Identification
Gene Expression
Peptide Hormones
Plant Development
Plant Protein
Abstract in English
Since the 90s, a new family of molecules of protein origin and with hormone characteristics has been studied in plants. This group of new molecules, collectively named peptide hormones, is involved in defense, reproduction, growth and development. RALF, one of these peptides, is ubiquitous in plants and is involved in plant development. In Arabidopsis there are 34 RALF-like genes (AtRALFs). Our group has shown that AtRALF1 is processed from a larger precursor by a subtilase. Arabidopsis has 56 subtilases, our goal is the identification of the specific subtilase that is responsible for the AtRALF1 processing. Prediction of subcelular localization and in silico gene expression analysis, both confirmed by RT-PCR expression analysis and chimeric proteins with green-fluorescent protein, allowed the reduction of the initial 56 candidates to only 7 subtilases. Crosses between knockout mutants or RNAi plants expressing low levels of subtilases with overexpressors of AtRALF1 identified the subtilases AtSBT6.1 (At5g19660) and AtSBT5.3 (At2g04160) as potentialy involved in the prepropeptide AtRALF1 processing.
 
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Publishing Date
2012-02-27
 
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