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Cloning and characterization of the gene from Arabidopsis thaliana PUMILIO

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Author(s):
Elaine Cristina Favaro
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
Defense date:
Examining board members:
Ronaldo Bento Quaggio; Carla Columbano de Oliveira; Mariana Cabral de Oliveira
Advisor: Ronaldo Bento Quaggio
Abstract

RNA-binding proteins often regulate the stability, localization, and translation of mRNAs by interaction with specific motifs in the 3-UTR. The PUMILIO protein from Drosophila was shown to control translation of specific mRNAs during development. PUMILIO homologs were found in several species and constitute the PUF family. The Arabidopsis thaliana sequencing project revealed four genes homolougs to PUMILIO. Three are situated in chromosome II (APUM-1, 2, and 3) that are nearly identical among themselves, and that contain a region that is 52% homologous to the PUMILIO. RNA-binding domain. The fourth is in chromosome IV (APUM-4), and shows low similarity to the other three. In this work, we characterized APUM-2 in further detail. Northern blots indicated that Apum-2 is expressed in shoot and root apices. Reporter transgenic plants were made that contained either APUM-2::GUS or Apum2::GFP constructs. Reporter gene expression confirmed that APUM-2 is active in shoot and root apices, specifically in the meristematic regions. Finally transgenic plants containing the 35S::APUM-2 construct were created. Constitutive APUM-2 expression resulted in plants with no apical dominance and a high number of leaves, stems and lateral roots. These results suggest that APUM-2 plays an important role during meristem development, possibly through a mRNA interaction. (AU)